20EM Bacterial Test I-chi
Kung I-chi《孔乙已》部分译文
The taverns in Lu Town are distinguishingly arranged compared with them in other towns. There are always, in the streets, carpenter's –square-shaped counters, in which prepared hot water is ready to warm the wine. Every day at noon and dusk, workers knocked off would spend four cents on a bottle of wine——that was the price twenty years ago, and it costs them ten now. They just stand in the street, against the counter, and enjoy their relaxing drinking time. And if you are ok with one cent more, you will get a dish of salty boiled bamboo shoot or beans flavored with aniseed as your appetizer. And over ten cents is needed if you even want some meat. However, the customers here are almost in short blouses, which kind of indicate their empty pockets. Only the ones with long gowns that show their wealth are qualified to step into the hall next to the tavern, sitting down and leisurely enjoy their food and wine.I have been a waiter in Xian Heng tavern since I was twelve. For the shopkeeper considered my appearance too foolish to serve the customers in long gowns inside, he just let me do some chores outside. Although it was easier to communicate with the short -blouses customers, I could be troubled by a fairly large of people who were always being wordy and inarticulate. They tended to watch the whole process from scooping out the rice wine from the jars to pouring the wine into the flagon, finally, staring the flagon in the hot water. Then, they were relieved, because they knew that it was difficult for me to add water in the wine under such close observation. So I was told to suspend the job just after a few days. Thanks to the broker, I was narrowly keptin Xian Heng tavern. But, instead, the shopkeeper shifted me to solely take charge of the wine-warming job.Since then I had to stay inside the counter all the day, focusing on my preoccupation. The job was a little bit monotonous despite my never-appeared dereliction of duty. Facing the fierce face of the shopkeeper and the discourtesy of the customers, I felt stiffness all the time. Only came Kung I-chi, I was offered the reason to laugh a little, which was so unforgettable.Kung I-chi was the only one who had his wine standing in the street while in long gown. He was a big man with jumbled grizzled beard. And there always were some scars on his wrinkled pale face. Long gown as he wore, the gown was dirty and shabby, which seemingly had been forgotten to get darned and even washed for over a decade. There were so many archaisms in his words that made the listeners difficult to fully understand him. Because his surname was Kung, so people just toke part of the elusive words on the copybook as his nickname. When Kung came to the tavern, all the people having their wine would look at him with smiles. And some of them would shout:―Kung I-chi! We see more scars on your face!‖No responding to them, he turned to the counter, and ordered:―Two bottles of wine and a dish of beans flavored with aniseed.‖Then he strugglingly pulled out all his treasure——nine cents.The people around Kung I-chi, on purposely, in a high-decibel voice, yelled to him again:―You must steal something again!‖―How can you frame innocent people without reasons?‖ Kung widened his eyes.―What innocence you are talking about? I personally saw you were hung up and beaten because you stole books from the Ho family!‖―Book stealing is not stealing! Stealing books is kind of scholar’s business, so can that be called stealing? ‖ Kung was blushing and exposing his blue veins on his forehead.And a string of incomprehensible words that quoted from some classics followed, making people around guffaw. So a merry atmosphere filled the tavern.I had heard something about Kung I-chi from people’s private talks. Kung used to be a scholar, but he failed the examination. Ignorance of making a living added to his poverty, and he was on the edge of begging. Fortunately, he mastered a good hand-writing, so he could struggle to live by some copy work. But he got a problem of his character, that is, he was addicted in drinking and was lazy. So after a few days he would disappear, taking books, paper, brushes and inkstone with him. After few times he did this, nobody let him do the copy work again. So doing some stealing is a natural and inevitable choice to make a living. But his character was ranked the top in our tavern. Namely, he never left debts. Although sometimes his name would be written on the blackboard because of his temporary lack of cash, he would clean his name by the laterpayment within a month.After half bottle wine, his reddened face gradually came to normal. And people around his asked him again:―Kung I-chi, are you really literate?‖Kung shot a glance at the asker with self-evident conceit.They asked continually:―How can you fail the examination the n?‖On hearing this, Kung’s gray face showed his gloominess and anxiety, murmuring all the archaisms, which were even more elusive. People at present guffawed at this scene, so a merry atmosphere filled the tavern again.The shopkeeper would not blame me if I echoed my laughter at this moment. Moreover, every time he met Kung I-chi, he was likely to ask him as usual to amuse people. Kung knew himself that it would be difficult to talk to adults, so he just turned his mouth to kids. Once he asked me:―Have you ever read some classics?‖I slightly nodded.―I see. So, let me test you. Do you know how to write the character hui in hui-xiang beans (beans flavored with aniseed)?‖I turned my back, thinking that a beggar was not qualified to test me.He waited for a long time, then, with earnest, he told me:―I know you can’t. I teach you, and you should remember this character, because it will be useful when you do the bookkeeping as a shopkeeper.‖I pondered that it would be so long for me to be a shopkeeper, and our shopkeeper never included beans flavored with aniseed in the bookkeeping.Thinking it funny but kind of tedious, I responded in an idle tone:―I don’t need your teaching. Isn't it the character hui with the grass radical?"Apparently he was happy about that, tapping the counter with his two long finger nails and nodding.―Yes, yes, you are right. Em…but do you know there are four forms for this same character?‖I finally lose my patience and left away in the pouts. And Kung just soaked his finger with wine, preparing to write them on the counter. My indifference made him sign and feel disappointed.Sometimes, the laughter attracted the kids in the neighborhood to join the merriment. Kung would hand out his beans one for each to the kids if they surrounded him. When they finished the only bean, they still stayed and stared at the dish. Kung, in a hurry, covered his beans with hand. Bent over, he said: ―L ittle, I only got little left.‖He straightened up and again peered at the dish, shaking his head, he said:―Little! Little! Indeed, not much!‖Then the kids dispersed with laughter.Kung I-chi owned such a power to make people happy; however, peoplestill live in this way without his presence.One day, two or three days before the Mid-Autumn Festival, the shopkeeper was dealing with accounting. After he took the blackboard, he suddenly shouted:―It has been so long since Kung I-chi came here last time, and he still owed me nineteen cents.‖I finally found his long-term absence was true.A customer responded:―How come he would turn up? His leg was beaten to fracture.‖―I see.‖ The shopkeeper said.―He still stole as usual. But this time, it was a huge mistake of him to steal from Mr. Ding, the provincial scholar! Nobody could steal in his house!‖―What happened then?‖―What happened then? First he was told to write a confession and a fierce beaten followed till midnight, and then his leg was in fracture.‖―Then?‖―Then they broke his leg.‖―And after that?‖―After? Who knows! He may be dead.‖The shopkeeper stopped his questions, and focused on the accounting.It was getting colder and colder after the Mid-Autumn Festival, which seemed like the beginning of winter. I had to wear padded coat and stay closeto the stove all day.One day afternoon, there is no business, so I sat with my eyes closed. All of a sudden, I heard something:―Warm a bowl of wine for me.‖The voice was low but familiar. I looked outside, nobody. And when I stood and looked, I found Kung I-chi sitting opposite to the threshold under the counter. His face was dark and skinny and was beyond image. He, with a shabby padded jacket, crossed his legs on a calceolaria hung on his neck with a rope.He saw me, and repeated:―Warm a bowl of wine for me‖The shopkeeper as well headed out to Kung, and said:―Are you Kung I-chi? You still owe me nineteen cents!‖He turned his face upwards, and shamefully said:―I… …I will settle it next time. I’ll pay you on cash today; the wine must be good. ‖The shopkeeper still talked to him with a smiley face as usual:―Kung I-chi, I heard you have stolen again!‖He didn’t try to argue on that this time. Instead, he briefly said:―No teasing me.‖―Teasing you? How come you were beaten to fracture if you didn’t steal?‖―Fell down… … fell… … I fell down.‖ He said in a low voice. And his eyeswere like to beg the shopkeeper to forget that.People gathered here, together with the shopkeeper, bursted into laughter.I warmed the wine, carried it outside, and put it on the threshold. He pulled out four cents from his pocket of the ragged jacket and laid them on my palm. Noticing his muddy hands, I guessed he must ―walk‖ here with them.A while later, after he finished his wine, he, sitting, slowly ―walked‖ away with his hands, accompanied with the talking and laughing of people.Since the n, I, again, haven’t seen Kung I-chi for such a long time. Time reached the near end of the year. The shopkeeper took down the blackboard and said:― Kung I-chi still owed me nineteen cents!‖ On the next Dragon Boat Festival, the shopkeeper repeated the same thing.But when the Mid-Autumn Festival came, he did not mention it. And another New Year came, and we still lose his news.I haven’t seen him ever since--maybe Kung I-chi is really dead.。
OECD 471 bacteria reverse mutation test 9747101E
471Adopted:21st July 1997 OECD GUIDELINE FOR TESTING OF CHEMICALSBacterial Reverse Mutation TestINTRODUCTION1.The bacterial reverse mutation test uses amino-acid requiring strains of Salmonella typhimurium and Escherichia coli to detect point mutations, which involve substitution, addition or deletion of one or a few DNA base pairs (1)(2)(3). The principle of this bacterial reverse mutationtest is that it detects mutations which revert mutations present in the test strains and restore the functional capability of the bacteria to synthesize an essential amino acid. The revertant bacteria are detected by their ability to grow in the absence of the amino acid required by the parent test strain.2.Point mutations are the cause of many human genetic diseases and there is substantial evidence that point mutations in oncogenes and tumour suppressor genes of somatic cells are involved in tumour formation in humans and experimental animals. The bacterial reverse mutationtest is rapid, inexpensive and relatively easy to perform. Many of the test strains have several features that make them more sensitive for the detection of mutations, including responsive DNA sequences at the reversion sites, increased cell permeability to large molecules and elimination of DNA repair systems or enhancement of error-prone DNA repair processes. The specificity of the test strains can provide some useful information on the types of mutations that are induced by genotoxic agents. A very large data base of results for a wide variety of structures is available for bacterial reverse mutation tests and well-established methodologies have been developed for testing chemicalswith different physico-chemical properties, including volatile compounds.3.Definitions used are set out in the Annex.INITIAL CONSIDERATIONS4.The bacterial reverse mutation test utilises prokaryotic cells, which differ from mammalian cells in such factors as uptake, metabolism, chromosome structure and DNA repair processes. Tests conducted in vitro generally require the use of an exogenous source of metabolic activation. In vitro metabolic activation systems cannot mimic entirely the mammalian in vivo conditions. The test therefore does not provide direct information on the mutagenic and carcinogenic potency of a substance in mammals.5.The bacterial reverse mutation test is commonly employed as an initial screen for genotoxic activity and, in particular, for point mutation-inducing activity. An extensive data base has demonstrated that many chemicals that are positive in this test also exhibit mutagenic activity in other tests. There are examples of mutagenic agents which are not detected by this test; reasons for these shortcomings can be ascribed to the specific nature of the endpoint detected, differences in metabolic activation, or differences in bioavailability. On the other hand, factors which enhance the sensitivity of the bacterial reverse mutation test can lead to an overestimation of mutagenic activity.471OECD/OCDE6.The bacterial reverse mutation test may not be appropriate for the evaluation of certainclasses of chemicals, for example highly bactericidal compounds (e.g. certain antibiotics) and those which are thought (or known) to interfere specifically with the mammalian cell replication system(e.g. some topoisomerase inhibitors and some nucleoside analogues). In such cases, mammalianmutation tests may be more appropriate.7.Although many compounds that are positive in this test are mammalian carcinogens, thecorrelation is not absolute. It is dependent on chemical class and there are carcinogens that are not detected by this test because they act through other, non-genotoxic mechanisms or mechanisms absent in bacterial cells.PRINCIPLE OF THE TEST METHOD8.Suspensions of bacterial cells are exposed to the test substance in the presence and in theabsence of an exogenous metabolic activation system. In the plate incorporation method, these suspensions are mixed with an overlay agar and plated immediately onto minimal medium. In the preincubation method, the treatment mixture is incubated and then mixed with an overlay agar before plating onto minimal medium. For both techniques, after two or three days of incubation, revertant colonies are counted and compared to the number of spontaneous revertant colonies on solvent control plates.9.Several procedures for performing the bacterial reverse mutation test have been described.Among those commonly used are the plate incorporation method (1)(2)(3)(4), the preincubation method (2)(3)(5)(6)(7)(8), the fluctuation method (9)(10), and the suspension method (11).Modifications for the testing of gases or vapours have been described (12).10.The procedures described in this guideline pertain primarily to the plate incorporation andpreincubation method s. Either of them is acceptable for conducting experiments both with and without metabolic activation. Some compounds may be detected more efficiently using the preincubation method. These compounds belong to chemical classes that include short chain aliphatic nitrosamines, divalent metals, aldehydes, azo-dyes and diazo compounds, pyrollizidine alkaloids, allyl compounds and nitro compounds (3). It is also recognised that certain classes of mutagens are not always detected using standard procedures such as the plate incorporation method or preincubation method. These should be regarded as "special cases" and it is strongly recommended that alternative procedures should be used for their detection. The following "special cases" could be identified (together with examples of procedures that could be used for their detection): azo-dyes and diazo compounds (3)(5)(6)(13), gases and volatile chemicals(12)(14)(15)(16), and glycosides (17)(18). A deviation from the standard procedure needs to bescientifically justified.DESCRIPTION OF THE METHODPreparationsBacteria11.Fresh cultures of bacteria should be grown up to the late exponential or early stationaryphase of growth (approximately 109 cells per ml). Cultures in late stationary phase should not be used. It is essential that the cultures used in the experiment contain a high titre of viable bacteria.The titre may be demonstrated either from historical control data on growth curves, or in each assay through the determination of viable cell numbers by a plating experiment.OECD/OCDE47112.The recommended culture temperature is 37°C.13.At least five strains of bacteria should be used. These should include four strains of S. typhimurium (TA1535; TA1537 or TA97a or TA97; TA98; and TA100) that have been shown to be reliable and reproducibly responsive between laboratories. These four S. typhimurium strains haveGC base pairs at the primary reversion site and it is known that they may not detect certain oxidising mutagens, cross-linking agents and hydrazines. Such substances may be detected by E.coli WP2 strains or S. typhimurium TA102 (19) which have an AT base pair at the primary reversion site. Therefore the recommended combination of strains is:1.S. typhimurium TA1535, and2.S. typhimurium TA1537 or TA97 or TA97a, and3.S. typhimurium TA98, and4.S. typhimurium TA100, and5. E. coli WP2 uvrA, or E. coli WP2 uvrA (pKM101), or S. typhimurium TA102.In order to detect cross-linking mutagens it may be preferable to include TA102 or to add a DNA repair-proficient strain of E.coli [e.g. E.coli WP2 or E.coli WP2 (pKM101).]14.Established procedures for stock culture preparation, marker verification and storage shouldbe used. The amino-acid requirement for growth should be demonstrated for each frozen stock culture preparation (histidine for S. typhimurium strains, and tryptophan for E. coli strains). Other phenotypic characteristics should be similarly checked, namely: the presence or absence of R-factor plasmids where appropriate [i.e. ampicillin resistance in strains TA98, TA100 and TA97a or TA97,WP2 uvrA and WP2 uvrA (pKM101), and ampicillin + tetracycline resistance in strain TA102]; the presence of characteristic mutations (i.e. rfa mutation in S. typhimurium through sensitivity to crystal violet, and uvrA mutation in E. coli or uvrB mutation in S. typhimurium, through sensitivity to ultra-violet light) (2)(3). The strains should also yield spontaneous revertant colony plate counts withinthe frequency ranges expected from the laboratory's historical control data and preferably within the range reported in the literature.Medium15.An appropriate minimal agar (e.g. containing Vogel-Bonner minimal medium E and glucose) and an overlay agar containing histidine and biotin or tryptophan, to allow for a few cell divisions, is used (1)(2)(9).Metabolic activation16.Bacteria should be exposed to the test substance both in the presence and absence of an appropriate metabolic activation system. The most commonly used system is a cofactor-supplemented post-mitochondrial fraction (S9) prepared from the livers of rodents treated with enzyme-inducing agents such as Aroclor 1254 (1)(2) or a combination of phenobarbitone and ß-naphthoflavone (18)(20)(21). The post-mitochondrial fraction is usually used at concentrations in the range from 5 to 30% v/v in the S9-mix. The choice and condition of a metabolic activation systemmay depend upon the class of chemical being tested. In some cases it may be appropriate to utilizemore than one concentration of post-mitochondrial fraction. For azo-dyes and diazo-compounds,using a reductive metabolic activation system may be more appropriate (6)(13).471OECD/OCDETest substance/Preparation17.Solid test substances should be dissolved or suspended in appropriate solvents or vehiclesand diluted if appropriate prior to treatment of the bacteria. Liquid test substances may be added directly to the test systems and/or diluted prior to treatment. Fresh preparations should be employed unless stability data demonstrate the acceptability of storage.Test conditionsSolvent/vehicle18.The solvent/vehicle should not be suspected of chemical reaction with the test substanceand should be compatible with the survival of the bacteria and the S9 activity (22). If other than well-known solvent/vehicles are used, their inclusion should be supported by data indicating their compatibility. It is recommended that wherever possible, the use of an aqueous solvent/vehicle be considered first. When testing water-unstable substances, the organic solvents used should be free of water.Exposure concentrations19.Amongst the criteria to be taken into consideration when determining the highest amount oftest substance to be used are cytotoxicity and solubility in the final treatment mixture. It may be useful to determine toxicity and insolubility in a preliminary experiment. Cytotoxicity may be detected by a reduction in the number of revertant colonies, a clearing or diminution of the background lawn, or the degree of survival of treated cultures. The cytotoxicity of a substance may be altered in the presence of metabolic activation systems. Insolubility should be assessed as precipitation in the final mixture under the actual test conditions and evident to the unaided eye. The recommended maximum test concentration for soluble non-cytotoxic substances is 5 mg/plate or5 µl/plate. For non-cytotoxic substances that are not soluble at 5 mg/plate or 5 µl/plate, one or moreconcentrations tested should be insoluble in the final treatment mixture. Test substances that are cytotoxic already below 5 mg/plate or 5 µl/plate should be tested up to a cytotoxic concentration.The precipitate should not interfere with the scoring.20.At least five different analysable concentrations of the test substance should be used withapproximately half log (i.e. √10) intervals between test points for an initial experiment. Smaller intervals may be appropriate when a concentration-response is being investigated.21.Testing above the concentration of 5 mg/plate or 5 µl/plate may be considered whenevaluating substances containing substantial amounts of potentially mutagenic impurities.Controls22.Concurrent strain-specific positive and negative (solvent or vehicle) controls, both with andwithout metabolic activation, should be included in each assay. Positive control concentrations that demonstrate the effective performance of each assay should be selected.23.For assays employing a metabolic activation system, the positive control referencesubstance(s) should be selected on the basis of the type of bacteria strains used. The following chemicals are examples of suitable positive controls for assays with metabolic activation:OECD/OCDE471Chemical and CAS No.9,10-Dimethylanthracene [CAS no. 781-43-1]7,12-Dimethylbenzanthracene [CAS no. 57-97-6]Congo Red [CAS no. 573-58-0] (for the reductive metabolic activation method)Benzo(a)pyrene [CAS no. 50-32-8]Cyclophosphamide (monohydrate) [CAS no. 50-18-0 (CAS no. 6055-19-2)]2-Aminoanthracene [CAS no. 613-13-8]2-Aminoanthracene should not be used as the sole indicator of the efficacy of the S9-mix. If 2-aminoanthracene is used, each batch of S9 should also be characterised with a mutagen that requires metabolic activation by microsomal enzymes, e.g., benzo(a)pyrene, dimethylbenzanthracene.24.For assays performed without metabolic activation system, examples of strain-specific positive controls are:Chemical and CAS No.Strain(a)Sodium azide [CAS no. 26628-22-8]TA1535 and TA100(b)2-Nitrofluorene [CAS no. 607-57-8]TA98(c)9-Aminoacridine [CAS no. 90-45-9]or ICR191 [CAS no. 17070-45-0]TA1537, TA97 and TA97a(d)Cumene hydroperoxide [CAS no. 80-15-9]TA102(e)Mitomycin C [CAS no. 50-07-7]WP2 uvrA and TA102(f)N-Ethyl-N-nitro-N-nitrosoguanidine [CAS no. 70-25-7] or4-nitroquinoline 1-oxide [CAS no. 56-57-5]WP2, WP2 uvrA and WP2 uvrA (pKM101)(g)Furylfuramide (AF-2) [CAS no. 3688-53-7]plasmid-containing strains25.Other appropriate positive control reference substances may be used. The use of chemical class-related positive control chemicals may be considered, when available.26.Negative controls, consisting of solvent or vehicle alone, without test substance, and otherwise treated in the same way as the treatment groups, should be included. In addition, untreated controls should also be used unless there are historical control data demonstrating that no deleterious or mutagenic effects are induced by the chosen solvent.471OECD/OCDEPROCEDURETreatment with test substance27.For the plate incorporation method (1)(2)(3)(4), without metabolic activation, usually 0.05ml or 0.1 ml of the test solutions, 0.1 ml of fresh bacterial culture (containing approximately 108 viable cells) and 0.5 ml of sterile buffer are mixed with 2.0 ml of overlay agar. For the assay with metabolic activation, usually 0.5 ml of metabolic activation mixture containing an adequate amount of post-mitochondrial fraction (in the range from 5 to 30% v/v in the metabolic activation mixture) are mixed with the overlay agar (2.0 ml), together with the bacteria and test substance/test solution.The contents of each tube are mixed and poured over the surface of a minimal agar plate. The overlay agar is allowed to solidify before incubation.28.For the preincubation method (2)(3)(5)(6) the test substance/test solution is preincubatedwith the test strain (containing approximately 108 viable cells) and sterile buffer or the metabolic activation system (0.5 ml) usually for 20 min. or more at 30°-37°C prior to mixing with the overlay agar and pouring onto the surface of a minimal agar plate. Usually, 0.05 or 0.1 ml of test substance/test solution, 0.1 ml of bacteria, and 0.5 ml of S9-mix or sterile buffer, are mixed with 2.0 ml of overlay agar. Tubes should be aerated during pre-incubation by using a shaker.29.For an adequate estimate of variation, triplicate plating should be used at each dose level.The use of duplicate plating is acceptable when scientifically justified. The occasional loss of a plate does not necessarily invalidate the assay.30.Gaseous or volatile substances should be tested by appropriate methods, such as in sealedvessels (12)(14)(15)(16).Incubation31.All plates in a given assay should be incubated at 37°C for 48-72 hours. After theincubation period, the number of revertant colonies per plate is counted.DATA AND REPORTINGTreatment of results32.Data should be presented as the number of revertant colonies per plate. The number ofrevertant colonies on both negative (solvent control, and untreated control if used) and positive control plates should also be given.33.Individual plate counts, the mean number of revertant colonies per plate and the standarddeviation should be presented for the test substance and positive and negative (untreated and/or solvent) controls.34.There is no requirement for verification of a clear positive response. Equivocal resultsshould be clarified by further testing preferably using a modification of experimental conditions.Negative results need to be confirmed on a case-by-case basis. In those cases where confirmation of negative results is not considered necessary, justification should be provided. Modification of study parameters to extend the range of conditions assessed should be considered in follow-up experiments.Study parameters that might be modified include the concentration spacing, the method of treatment (plate incorporation or liquid preincubation), and metabolic activation conditions.OECD/OCDE471 Evaluation and interpretation of results35.There are several criteria for determining a positive result, such as a concentration-related increase over the range tested and/or a reproducible increase at one or more concentrations in the number of revertant colonies per plate in at least one strain with or without metabolic activation system (23). Biological relevance of the results should be considered first. Statistical methods maybe used as an aid in evaluating the test results (24). However, statistical significance should not bethe only determining factor for a positive response.36. A test substance for which the results do not meet the above criteria is considered non-mutagenic in this test37.Although most experiments will give clearly positive or negative results, in rare cases thedata set will preclude making a definite judgement about the activity of the test substance. Resultsmay remain equivocal or questionable regardless of the number of times the experiment is repeated.38.Positive results from the bacterial reverse mutation test indicate that a substance inducespoint mutations by base substitutions or frameshifts in the genome of either Salmonella typhimuriumand/or Escherichia coli. Negative results indicate that under the test conditions, the test substance isnot mutagenic in the tested species.Test report39.The test report must include the following information:Test substance:-identification data and CAS no., if known;-physical nature and purity;-physicochemical properties relevant to the conduct of the study;-stability of the test substance, if known.Solvent/Vehicle:-justification for choice of solvent/vehicle;-solubility and stability of the test substance in solvent/vehicle, if known.Strains:-strains used;-number of cells per culture;-strain characteristics.Test conditions:-amount of test substance per plate (mg/plate or µg/plate) with rationale for selection of dose and number of plates per concentration;-media used;-type and composition of metabolic activation system, including acceptability criteria;-treatment procedures.471OECD/OCDEResults:-signs of toxicity;-signs of precipitation;-individual plate counts;-the mean number of revertant colonies per plate and standard deviation;-dose-response relationship, where possible;-statistical analyses, if any;-concurrent negative (solvent/vehicle) and positive control data, with ranges, means and standard deviations;-historical negative (solvent/vehicle) and positive control data, with e.g. ranges, means and standard deviations.Discussion of the results.Conclusion.LITERATURE(1)Ames, B.N., McCann, J. and Yamasaki, E. (1975). Methods for Detecting Carcinogens andMutagens with the Salmonella/Mammalian-Microsome Mutagenicity Test. Mutation Res., 31,347-364.(2)Maron, D.M. and Ames, B.N. (1983). Revised Methods for the Salmonella Mutagenicity Test.Mutation Res., 113, 173-215.(3)Gatehouse, D., Haworth, S., Cebula, T., Gocke, E., Kier, L., Matsushima, T., Melcion, C.,Nohmi, T., Venitt, S. and Zeiger, E. (1994). Recommendations for the Performance ofBacterial Mutation Assays. Mutation Res., 312, 217-233.(4)Kier, L.D., Brusick D.J., Auletta, A.E., Von Halle, E.S., Brown, M.M., Simmon, V.F., Dunkel,V., McCann, J., Mortelmans, K., Prival, M., Rao, T.K. and Ray V. (1986). The SalmonellaTyphimurium/Mammalian Microsomal Assay: A Report of the U.S. Environmental ProtectionAgency Gene-tox Program. Mutation Res., 168, 69-240.(5)Yahagi, T., Degawa, M., Seino, Y.Y., Matsushima, T., Nagao, M., Sugimura, T. andHashimoto, Y. (1975). Mutagenicity of Carcinogen Azo Dyes and their Derivatives. CancerLetters, 1, 91-96.(6)Matsushima, M., Sugimura, T., Nagao, M., Yahagi, T., Shirai, A., and Sawamura, M. (1980).Factors Modulating Mutagenicity Microbial Tests. In: Short-term Test Systems for DetectingCarcinogens. Ed. Norpoth K.H. and Garner, R.C., Springer, Berlin-Heidelberg-New York. pp.273-285.(7)Gatehouse, D.G., Rowland, I.R., Wilcox, P., Callender, R.D. and Foster, R. (1990). BacterialMutation Assays. In: Basic Mutagenicity Tests: UKEMS Part 1 Revised. Ed. D.J. KirklandCambridge University Press, pp. 13-61.(8)Aeschbacher, H.U., Wolleb, U. and Porchet, L. (1987). Liquid Preincubation Mutagenicity Testfor Foods. J. Food Safety, 8, 167-177.OECD/OCDE471 (9)Green, M. H. L., Muriel, W. J. and Bridges, B.A. (1976). Use of a simplified fluctuation test todetect low levels of mutagens. Mutation Res., 38, 33-42.(10)Hubbard, S.A., Green, M.H.L., Gatehouse, D., and J.W. Bridges (1984). The Fluctuation Test inBacteria. In: Handbook of Mutagenicity Test Procedures. 2nd Edition. Ed. Kilbey, B.J.,Legator , M.,Nichols, W. and Ramel C., Elsevier, Amsterdam-New York-Oxford,pp. 141-161. (11)Thompson, E.D. and Melampy, P.J. (1981). An Examination of the Quantitative SuspensionAssay for Mutagenesis with Strains of Salmonella typhimurium. Environmental Mutagenesis,3, 453-465.(12)Araki, A., Noguchi, T., Kato, F. and T. Matsushima (1994). Improved Method forMutagenicity Testing of Gaseous Compounds by Using a Gas Sampling Bag. Mutation Res.,307, 335-344.(13)Prival, M.J., Bell, S.J., Mitchell, V.D., Reipert, M.D. and Vaughn, V.L. (1984). Mutagenicityof Benzidine and Benzidine-Congener Dyes and Selected Monoazo Dyes in a ModifiedSalmonella Assay. Mutation Res., 136, 33-47.(14)Zeiger, E., Anderson, B. E., Haworth, S, Lawlor, T. and Mortelmans, K. (1992). SalmonellaMutagenicity Tests. V. Results from the Testing of 311 Chemicals. Environ. Mol. Mutagen., 19,2-141.(15)Simmon, V., Kauhanen, K. and Tardiff, R.G. (1977). Mutagenic Activity of ChemicalsIdentified in Drinking Water. In Progress in Genetic Toxicology, D. Scott, B. Bridges and F.Sobels (Eds.)., Elsevier, Amsterdam, pp. 249-258.(16)Hughes, T.J., Simmons, D.M., Monteith, I.G. and Claxton, L.D. (1987). VaporizationTechnique to Measure Mutagenic Activity of Volatile Organic Chemicals in theAmes/Salmonella Assay. Environmental Mutagenesis, 9, 421-441.(17)Matsushima, T., Matsumoto, A., Shirai, M., Sawamura, M. and Sugimura, T. (1979).Mutagenicity of the Naturally Occurring Carcinogen Cycasin and Synthetic MethylazoxyMethane Conjugates in Salmonella typhimurium. Cancer Res., 39, 3780-3782.(18)Tamura, G., Gold, C., Ferro-Luzzi, A. and Ames. B.N. (1980). Fecalase: A Model forActivation of Dietary Glycosides to Mutagens by Intestinal Flora. Proc. Natl. Acad. Sci. USA,77, 4961-4965.(19)Wilcox, P., Naidoo, A., Wedd, D. J. and Gatehouse, D. G. (1990). Comparison of Salmonellatyphimurium TA 102 with Escherichia coli WP2 Tester strains. Mutagenesis, 5, 285-291.(20)Matsushima, T., Sawamura, M., Hara, K. and Sugimura, T. (1976). A Safe Substitute forPolychlorinated Biphenyls as an Inducer of Metabolic Activation Systems. In: "In vitroMetabolic Activation in Mutagenesis Testing", Eds F.J. de Serres et al. Elsevier, North Holland,pp. 85-88.(21)Elliott, B.M., Combes, R.D., Elcombe, C.R., Gatehouse, D.G., Gibson, G.G., Mackay, J.M. andWolf, R.C. (1992). Alternatives to Aroclor 1254-induced S9 in in vitro Genotoxicity Assays.Mutagenesis, 7, 175-177.(22)Maron, D., Katzenellenbogen, J., and Ames, B.N. (1981). Compatibility of Organic Solventswith the Salmonella/Microsome Test. Mutation Res., 88, 343-350.471OECD/OCDE(23)Claxton, L.D., Allen, J., Auletta, A., Mortelmans, K., Nestmann, E., and Zeiger, E., (1987).Guide for the Salmonella typhimurium/Mammalian Microsome Tests for BacterialMutagenicity. Mutation Res., 189, 83-91.(24)Mahon, G.A.T., Green, M.H.L., Middleton, B., Mitchell, I., Robinson, W.D. and Tweats, D.J.(1989). Analysis of Data from Microbial Colony Assays. In: UKEMS Sub-Committee onGuidelines for Mutagenicity Testing Part II. Statistical Evaluation of Mutagenicity Test Data.Ed. Kirkland, D.J., Cambridge University Press, pp. 28-65.OECD/OCDE471ANNEXDEFINITIONSA reverse mutation test in either Salmonella typhimurium or Escherichia coli detects mutation in an amino-acid requiring strain (histidine or tryptophan, respectively) to produce a strain independent of an outside supply of amino-acid.Base pair substitution mutagens are agents that cause a base change in DNA. In a reversion test this change may occur at the site of the original mutation, or at a second site in the bacterial genome.Frameshift mutagens are agents that cause the addition or deletion of one or more base pairs in the DNA, thus changing the reading frame in the RNA11/11。
食品微生物自动化仪器检测
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01
第一节 全自动微生物鉴定仪器
02
第二节 全自动微生物总数和大肠杆菌快速测定仪器
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第三节 其他快速测定仪器
第十五章 食品微生物自动化仪器检测
检测和鉴定细菌的标准方法是什么？
ATB Expression 细菌鉴定智能系统
自动菌落计数系统
全自动酶联荧光免疫分析系统（mini VIDAS）
应用电阻抗技术的全自动微生物监测系统 —— BACTOMETER
API—细菌鉴定系统
第三节 其他快速测定仪器
第三节 其他快速测定仪器
自动菌落计数系统
第三节 其他快速测定仪器
二、应用电阻抗技术的全自动微生物监测系统 —— BACTOMETER P351 电阻抗检测法是应用于细菌检测的一项电化学技术，５０年代美国开始出现以此为原理的自动化仪器。７０年代～８０年代欧洲也陆续开发出类似的仪器。1998年电阻抗分析法成为除平板分析法外其他微生物检测法中首先被German standardization institution接受的微生物测定方法。
主要操作程序
用光电比浊仪配制菌悬液，将待检菌液浓度调节至各试条要求的范围。
将试条置入普通培养箱35℃培养4或24小时。
将试条置入读数器读数，并打印报告。 报告内容包括细菌鉴定、药敏试验S、I、R（部分试条含MIC）的结果，有丰富的专家系统信息，严格控制报告质量。
ATB Expression 细菌鉴定智能系统
第三节 其他快速测定仪器
全自动酶联荧光免疫分析系统（mini VIDAS）
Mini VIDAS performs assays using ELFA (Enzyme Linked Fluorescent Assay) technology.
大肠埃希氏菌产ESBLs和AmpC酶的检测
出率为 ６ ０ ％ 。结论 ．３
笔者所在医院大肠埃 希氏菌主要产 Ｅ Ｂ ｓ ， Ｓ Ｌ 酶 其次是 Ａ ｐ ｍ Ｃ酶 ， 酶菌株临床经验 用药应首 产
选碳青霉烯 类， 其次是 Ｂ一内酰胺类抗菌素 ＋酶抑制剂 、 头霉素类 ， 及其 它敏感抗菌素 。
【 关键词 】 大肠埃希氏菌 ； 超广谱 ｐ 一内酰胺酶 （ Ｓ Ｌ ） 头孢菌 素酶（ Ｍ Ｃ ＥＢｓ； Ａ Ｐ）
１ ５例 ， 列 腺 液 ６例 ， 汁 ７例 ， 髓 １例 。 前 胆 骨 １２ 细 菌 鉴 定 ． 进行鉴定。 １２ １ Ｅ Ｂ ． ． Ｓ Ｌ确 证 实 验 以 大 肠 埃 希 氏 菌 （ Ｔ Ｃ ５２ ） 肺 Ａ Ｃ ２９２ 、 采 用 法 国梅 里 埃 公 司 Ｖ Ｋ Ｉ Ｅ一２细 菌 分 析 仪
菌 环 直 径 ≥５ｍ 即判 断 为 产超 广谱 Ｂ一内 酰 胺 酶 。 ｍ，
１２ ２ Ａ Ｃ酶 表 型 筛 选 ． ． ｍｐ
行 药 敏 试 验 ， 进 行 表 型筛 选 。判 断 标 准 按 照 Ｎ Ｃ Ｓ推 荐 的 并 ＣＬ
１ １ 菌株来源 ．
３ ５株大肠埃希 氏菌菌来 自 ２０ ６ ０ ６年 １月 ～
２０ ０ ８年 １ ２月笔者所 在 医院分 离 的 阳性标 本 ， 中痰 液 １０ 其 ３ 例， 气管刷取 物 ５例 ， 尿液 ９ ， ４例 血液 １ ， ７例 胸水 ２例 ， 腹水 ２ 例， 脑脊液 １ ， 例 分泌物 ７ ２例 ， 引流物 ８例 ， 穿刺液 ５例 ， 脓液
埃希 氏菌 中有 ９ ８株 单 产 Ｅ Ｂ ｓ 检 出 率 ２． ％ ， ＳＬ， ６ ８ ６株单 产 Ａ ｐ ｍ Ｃ酶 ， 检出率 １６ ，２株 同时 产 Ｅ Ｂ ｓ和 Ａ Ｃ酶 ， ．％ ２ ＳＬ ｍｐ 占
免疫胶体金技术快速检测大肠杆菌O157和志贺毒素
上海交通大学硕士学位论文免疫胶体金技术快速检测大肠杆菌O157和志贺毒素姓名：高成秀申请学位级别：硕士专业：预防兽医学指导教师：严亚贤;孙建和20080101免疫胶体金技术快速检测大肠杆菌O157和志贺毒素摘要大肠杆菌O157是产志贺毒素大肠杆菌（Shiga-producing Escherichia coli, STEC）的主要血清型，可以引起人和动物的腹泻、出血性结肠炎、溶血性尿毒综合征、血栓性血小板减少性紫癜。
志贺毒素（Shiga toxin，Stxs）是大肠杆菌 O157的主要毒力因子之一，有两个生物型：Stx1和Stx2，只产生Stx2的菌株毒力比只分泌Stx1和两者同时分泌的菌株毒力都强。
Stx2致病剂量极低，且分泌胞外，Stx2的检测将直接有助于对分离菌株致病力强弱的判定，提高对高致病性大肠杆菌O157菌株的检出。
因此本课题研究大肠杆菌O157和Stx2的胶体金层析法(Ｇｏｌｄ Ｉｍｍｕｎｏｃｈｒｏｍａｔｏｇｒａｐｈｙ Ａｓｓａｙ， ＧＩＣＡ)，进行高致病性大肠杆菌O157菌株检测和鉴定，以便更好的预防和控制大肠杆菌O157所致的人和动物的疾病。
将本实验室构建的重组Stx2B亚单位的质粒pGEX-Stx2B转化入大肠杆菌BL21，然后采用PCR和酶切进行鉴定，将阳性转化菌进行重组蛋白的表达、鉴定、纯化，用纯化的融合蛋白多次免疫新西兰大白兔制备抗重组Stx2B的多克隆抗体。
同时用大肠杆菌O157 ATCC43889全菌体抗原多次免疫新西兰大白兔，制备抗大肠杆菌O157多克隆抗体。
用硫酸胺法、辛酸-硫酸胺法、Protein G亲和层析3种方法分别纯化制备的多克隆抗体，结果表明：Protein G法纯化的抗重组Stx2B IgG纯度较其前两者纯，其金标多抗的稳定性较好，辛酸-硫酸胺法纯化的抗大肠杆菌O157 IgG的活性较Protein G纯化的好，其金标多抗的稳定性较好，因此本试验采用Protein G法纯化抗Stx2B 血清，用辛酸-硫酸胺法纯化抗大肠杆菌O157血清。
API20E生化鉴定条在致病菌检验中的应用
API20E生化鉴定条在致病菌检验中的应用摘要】目的提高API20E鉴定系统在肠道致病菌鉴定中的应用。
方法应用API20E和国产生化鉴定管对13株阳性菌株（4株致病性大肠埃希氏菌、7株志贺氏菌、2株甲型副伤寒菌）进行鉴定比较，并经血清学凝集验证。
结果表明API20E鉴定系统方法程序简化，检测结果准确可靠，对致病菌鉴定有现实的指导意义，可作为常规致病菌检测方法。
【关键词】API20E生化鉴定条、生化鉴定管、沙门氏菌、志贺氏菌【中图分类号】R446 【文献标识码】A 【文章编号】2095-1752（2014）30-0305-02近年来新的生化鉴定系统不断推出，以传统的试验方法来鉴定致病菌已不能适应准确、快速诊断的需求。
API20E生化鉴定系统是由法国生物梅里埃公司生产细菌数值分类鉴定系统，它的鉴定能力强，数据库不断更新和补充完善，已被许多国家和组织列入标准方法。
现在就API20E生化鉴定系统与国产生化鉴定官的鉴定效果进行比较对比，报告如下：1、样品、材料和检测依据1.1样品来源：本地区历年检测出的阳性菌株13株（4株致病性大肠埃希氏菌、7株志贺氏菌、2株甲型副伤寒菌）1.2、材料：API20E生化反应试剂条（法国生物梅里埃公司生产），批号：1002230950；API-Plus鉴定软件；国产细菌生化鉴定管（北京路桥公司生产），批号：20140218；志贺氏菌属诊断血清22种（宁波天润生物药业公司生产），批号：20140201；肠道致病性大肠埃希氏菌诊断学清15种（宁波天润生物药业公司生产）批号：20140201；沙门氏菌属诊断血清30种（宁波天润生物药业公司生产），批号：20140201。
1.3、检验依据：沙门氏菌检验方法：GB/T4789.4-2010、志贺氏菌检验方法：GB/T4789.5-2012、致泻性大肠埃希氏菌检验方法：GB/T4789.6-2003。
API20E生化反应试剂条和国产细菌生化鉴定管按厂家操作说明书进行操作。
梅里埃微生物药敏鉴定分析仪操作规程
梅里埃ATB微生物药敏鉴定分析仪操作规程1 主要技术指标梅里埃ATB微生物鉴定／药敏分析系统融合了自动化、电脑化及微生物微量生化反应测试方法，可同时进行微生物分析鉴定（ID）与药敏（MIC）检测，使细菌鉴定／药敏分析规范化、标准化、现代化。
1.1完善的分析系统：微生物鉴定分析系统，微生物药敏分析系统统计分析／院内感染监测专家系统联网功能1.2药物种类：呋喃类，磺胺类，喹诺酮类，青霉素类大环内酯类，氨基糖甙类，头孢菌素类1.2鉴定范围：肠杆菌科，非发酵菌，葡萄球菌属，真菌，微球菌属，链球菌属，肠球菌属，弧菌属1.3院内感染检测环境空气，物体表面，医疗用品，消毒剂及保存液，血液透析2 适用范围梅里埃ATB微生物分析系统(包括微生物鉴定和药敏分析两大功能)；是对己分离培养出来的细菌、真菌等微生物进行种、属鉴定，并同时测定该菌对各种抗菌药物敏感/耐药程度的专用设备。
3 工作原理自1880年郭霍(Koch)发明固体培养基，对细菌的研究技术有了重大的突破之后，一个多世纪以来，人类对细菌的研究和认识已经得到了极大的成就，对细菌分类的理论根据和方法已趋成熟，命名已经统一，对繁浩的各种细菌众多的生物学、物理学特性己基本明确，从20世纪初开始，形成了利用各种细菌之间生物学、物理学特性的差异或不同，来区别和鉴定细菌种类的方法，不断丰富发展，沿用至今，成为细菌鉴定的常规方法。
但由于细菌种类繁多，生物学性状错综复杂，实验操作繁琐、费时，不仅初学者不易掌握，有经验者也常感困惑。
20世纪70年代，由于电子计算机的发展，有人首先利用信息编码来签定细菌，将复杂的各种细菌的生物学性状建立数据库，被检菌的各种生物性状测出后，输入电脑与贮存信息相比较即刻可判断未知菌的种属。
八十年代初己设计出各种多项生物学试验的套装组合，与电脑贮存的项目内容相对应，即细菌的编码鉴定法，很快在全球得到了重视，迅速推广应用。
随后，在上述编码鉴定的基础上，又开发研究了各种类型的半自动和全自动的细菌鉴定药敏分析仪器。
梅里埃全自动微生物鉴定仪参数
梅里埃全自动微生物鉴定仪参数设备名称：全自动微生物鉴定及药敏分析系统一、具体用途：对食品，环境中的微生物进行快速，全自动的鉴定及药物敏感性测试。
二、技术参数与性能要求：1. 系统可同时处理》30个标本，系统具有扩容功能，至少可以两台联机；2. 分析组件可对环境中和食品中的细菌进行全自动鉴定，种类包括革兰阴性菌、革兰阳性球菌、革兰氏阳性杆菌、酵母样真菌、假丝酵母类真菌、苛养菌、厌氧菌及棒状杆菌等的鉴定；3. ★分析组件可对芽孢杆菌进行全自动鉴定；4. ★大于500种可鉴定细菌，鉴定结果通过美国FDA认证，细菌鉴定采用GB推荐生化鉴定显色法，药敏检测采用比浊法，并且鉴定方法原理可在GB4789中查询（提供具体细菌库）；5. ^分析组件可自动进行革兰阴性菌、革兰阳性菌、酵母样真菌、肺炎链球菌等药敏试验，以上所有药敏试验均得到美国FDA批准用于临床应用（提供FDA 证明资料）；6. ★在对标本的鉴定及药敏试验过程中，无需添加任何额外附加试剂；7. 快速全自动对细菌进行鉴定和药敏试验，采用实时检测系统，系统每隔15分钟对试剂卡进行一次扫描读数，一旦确认结果，可马上出报告；& ★细菌最快鉴定时间V 4个小时，平均鉴定时间不超过5小时；9?最快药敏实验时间5小时，平均药敏实验时间不大于6小时；10. ★系统可同时进行鉴定和药敏实验，并且可同时上机的鉴定试剂卡种类不少于4种，可同时上机的药敏试剂卡的种类不少于6种；11. ★系统自动填充悬浮液至试剂卡，自动密封拭卡，并自动将拭卡装载于设备内置读数系统/孵育系统，测试结束时可自动丢弃拭卡，操作都在仪器内部自动进行，不需要额外设备；12. 卡片填充菌液后为封闭式卡片，不会造成污染；13. ★鉴定卡和药敏卡必须独立包装；14. 鉴定卡应至少提供3种不同试剂的SFDA注册证；15. 药敏卡应至少提供5种不同试剂的SFDA注册证；16. 测试完成后，经分析软件分析后得出结果并可自动打印报告，并保存结果；17. 具备中文报告软件系统；18?双向联网软件，可传输报告结果；19?具有三重售后服务保证体系（国内有分支机构、本地有生产厂家办事处、销售商有专职工程师），必须提供终身售后服务支持;20.售后服务时效：有仪器故障，厂方能在4小时作出响应。
美国药典微生物限度检测
61微生物限度检测（MICROBIAL LIMIT TESTS）此章提供方法来检测可能存在的好氧微生物其他制药过程中可能出现的微生物的数量，包括原材料和成品中的。
如果经过验证确认可以得到相同或更好的检测结论，也允许采用自动化的检测方法。
在样品检测过程中须进行无菌操作。
若无特别说明，则“培养（incubate）”一词指在30—35℃的培养箱内培养24至48小时；“生长（growth）”一词用于专门的判定，说明“存在和可能存在活的微生物”。
准备实验 (Preparatory Testing)本章涉及实验结果的有效性取决于：提供的被检测样品本身在实验条件下，被充分证明不会抑制可能存在的微生物的生长。
因此，在准备样品时，需要正规的实验操作和符合要求的实验条件，接种稀释样品到含有以下（微生物）培养物的培养基：金黄色（奥里斯）葡萄球菌（Staphylococcus aureus）,大肠埃希氏菌（Escherichia coli）, 铜绿假单胞菌(Pseudomonas aeruginosa), 和沙门氏菌（Salmonella）。
方法如下：将用肉汤培养基培养24小时后的（微生物）不小于10-3稀释的微生物培养物，加1 ml（微生物）培养液到磷酸(盐)缓冲液(pH 7.2)，液体大豆酪蛋白消化物培养基（Fluid Soybean-Casein Digest Medium），或者液体乳糖培养基（Fluid Lactose Medium）。
相应培养基培养失败则需要采取以下方法更改检测程序：（1）增加稀释液体积，检测样品加入量仍维持不变；或者（2）中和一定数量的干扰因子；或者（3）结合（1）、（2）得出适当条件，使接种物得以生长。
以下是一些物质的成分和浓度，该物质及浓度可用于加入培养基、阻止物质发挥抑菌作用：大豆卵磷脂（soy lecithin, 0.5%）或者聚山梨醇酯20（polysorbate 20, 4.0%）。
《医学统计概论》第7章卡方检验Chi-square test
(3) 当n＜40或有T＜1时，用Fisher’s exact probability。
7.2 配对四格表资料的χ2检验
配对设计包括：（1）同一批样品用两种不同的处理方法；（2）观察 对象根据配对条件配成对子，同一对子内不同的个体分别接受不同的处理； （3）在病因和危险因素的研究中，将病人和对照按配对条件配成对子， 研究是否存在某种病因或危险因素。
表7-1 两组降低颅内压有效率的比较（P137）
组别
试验组 对照组 合计
有效
99 75 174
无效
5 21 26
合计
104 96 200
有效率(%)
95.20 (p1) 78.13 (p2) 87.00 (pc)
实际频数A (actual frequency) 理论频数T (theoretical frequency)
,
1
因为有一格1＜T＜5，且n＞40时，所以应用连续性校
正χ2检验。
四、精确概率法(Fisher’s exact probability)
在无效假设成立的前提下且周边合计固定时，产生任意 一个四格表(i)的概率Pi 服从于超几何分布，其计算式为：
a b!c d !a c!b d !
Pi
a!b!c!d !n!
药物治疗组 164
18
182
外用膏药组 118
26
144
4.59
>0.0125 (NS)
合计
282
44
326
二、各实验组与同一对照组比 关键是检验水平的校正
'
2k 1
自学
7.6 双向有序分组资料的线性趋势检验
抗菌药物最小抑菌浓度的测定
抗菌药物最小抑菌浓度的测定一、概念：最小抑菌浓度（minimum inhibitory concentration, MIC）：在特定环境下孵育24小时，可抑制某种微生物出现明显增长的最低药物浓度即最小抑菌浓度，用于定量测定体外抗菌活性。
二、实验目的:通过采用常量稀释法, 检测几种抗菌药物对—--—菌株的最小抑菌浓度（MIC) ，对临床诊断用药有指导作用.三、仪器和试剂：四、检验方法:液体稀释法、琼脂稀释法、E试验（E test) 液体稀释法操作步骤：1.。
抗菌药物原液制备抗菌药物贮存液浓度不应低于1000μg/ml（如1280μg/ml)或10倍于最高测定浓度.溶解度低的抗菌药物可稍低于上述浓度.所需抗菌药物溶液量或粉剂量可公式进行计算.例如：需配制100 ml浓度为1280μg/ml的抗生素贮存液，所用抗生素为粉剂，其药物的有效力为750μg/mg。
用分析天平精确称取抗生素粉剂的量为182。
6 mg。
根据公式计算所需稀释剂用量为：（182。
6 mg×750μg/ml）/1280μg/ml=107。
0ml，然后将182.6 mg抗生素粉剂溶解于107.0ml稀释剂中。
制备抗菌药物贮存液所用的溶剂和稀释剂见表1.配制好的抗菌药物贮存液应贮存于—20℃环境，并注意抗菌药物的保存期限。
表1 稀释法中常用的抗菌药物容积稀释法2.药敏试验用抗菌药物浓度范围根据NCCLS抗菌药物敏感性试验操作标准,药物浓度范围应包含耐药、中介和敏感分界点值，特殊情况例外。
3.培养基NCCLS推荐使用Mueller-Hinton（MH）肉汤，pH7。
2~7。
4。
需氧菌及兼性厌氧菌在此培养基中生长良好。
在测试葡萄球菌对苯唑西林的敏感性时，应在肉汤中加入2%（W/V)氯化钠，按制造厂家的要求配制需要量的MH肉汤。
嗜血杆菌属菌使用HTM肉汤,肺炎链球菌和其它链球菌使用含2％～5％溶解马血的MH肉汤。
4。
接种物的制备有2种方法配制接种物，一是细菌生长方法，用接种环挑取形态相似待检菌落3-5个,接种于4-5ml的水解酪蛋白（MH)肉汤中,35℃孵育2-6h。
API20E 肠道菌试剂鉴定条详解
法国梅里埃公司API20E 肠道菌试剂鉴定条上面就是API20E的鉴定试剂条，也就是用来鉴定肠道细菌之用途的。
一共是20种生化反应再加手工操作的氧化酶试验（ARA后面的OX）。
非常赞叹梅里埃公司的厉害，能把非常复杂的生化反应做成那么小的试剂条，而且还做了细菌生化反应数据库，好庞大的工作量和好精细的工作…………下面我来分别介绍一下这20种生化反应原理以及用途：复杂但是很有趣味……1）ONPG试验翻译成中文是：邻硝基苯-半乳糖苷酶试验。
用来检测细菌是否发酵乳糖。
原理是因为乳糖发酵过程中需要乳糖通透酶和β－半乳糖苷酶才能快速分解，但是有些细菌没有乳糖通透酶，只有半乳糖苷酶，所以会导致迟缓发酵。
由此可得，如果某种细菌能够发酵乳糖，那么一定会产生β－半乳糖苷酶。
ONPG试验是将ONPG作为底物，如果细菌产生β－半乳糖苷酶，那么会水解ONPG为半乳糖和黄色的邻-硝基苯酚（ONP），培养液会变成黄色。
因此试验结果为阳性。
2）ADH 精氨酸双水解酶试验:某些细菌能产生精氨酸水解酶，先水解精氨酸生成瓜氨酸和氨，再水解瓜氨酸生成鸟氨酸、氨和二氧化碳。
因此使培养基呈碱性。
用于阴沟肠杆菌（阳性）和产气肠杆菌（阴性）的鉴别。
（氨基酸分解过程是在厌氧环境中进行的，所以要加盖石蜡油造成厌氧环境）。
3）LDC 赖氨酸脱羧酶试验：某些细菌能产生赖氨酸脱羧酶，可以将赖氨酸脱羧生成胺和二氧化碳，但是碱性更强，使培养基呈紫色（溴甲酚紫指示剂）。
用来鉴别用于产气肠杆菌（阳性）与阴沟肠杆菌（阴性）。
4）ODC 鸟氨酸脱羧酶试验：某些细菌产生鸟氨酸脱羧酶，将鸟氨酸脱羧生成胺和二氧化碳，是培养基呈碱性。
用来鉴别鸟氨酸用于阴沟肠杆菌（阳性）和克雷伯菌（阴性）。
5）CIT 柠檬酸盐（又称枸橼酸盐）利用实验：当柠檬酸盐作为唯一的碳源，某些细菌能利用此碳源而分解柠檬酸盐产生碳酸盐，而使培养基呈碱性由浅绿色变成深蓝色。
肠杆菌科中：沙门菌属、克雷伯菌属、粘质和液化沙雷菌及某些变形杆菌以及枸橼酸杆菌阳性；而埃希菌属、志贺菌属、爱德华菌属和耶尔森菌属均为阴性。
大肠埃希氏菌(内容详细)
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6.3平板菌落数的选择
数在10 CFU～100 CFU之间的 平板，暗室 360nm～ 366nm 波 长紫外灯照
射下，计数
平板上发浅
蓝色荧光的 菌落。
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6.2 操作步骤
检验时用已知MUG阳性菌株（如大肠埃希 氏菌ATCC25922）和产气肠杆菌（如
ATCC 13048）做阳性和阴性对照。
的样品匀液，每个稀释度接种2个无菌平皿， 每皿1 mL。同时取1 mL稀释液加入无菌 平皿做空白对照。
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6.2 操作步骤
6.2.2.2 将10 mL～15 mL冷至 45℃±0.5℃的结晶紫中性红胆盐琼脂 （VRBA）倾注于每个平皿中。小心旋转 平皿，将培养基与样品匀液充分混匀。
6.2.2.2 …待琼脂凝固后，再加3mL～4 mL VRBA-MUG覆盖平板表层。凝固后 翻转平板，36 ℃±1 ℃培养18 h～24 h。
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1 范围
本标准规定了食品中大肠埃希氏菌 （Escherichia coli）计数的方法。
本标准适用于食品中大肠埃希氏菌的计数， 其中大肠埃希氏菌平板计数法（第二法） 不适用于贝类产品。
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2 术语和定义
2.1 大肠埃希氏菌 Escherichia coli 大肠杆菌广泛存在于人和温血动物的肠道 中，能够在44.5℃发酵乳糖产酸产气， IMViC（靛基质、甲基红、VP试验、柠檬 酸盐）生化试验为＋＋――或－＋――的 革兰氏阴性杆菌。以此作为粪便污染指标 来评价食品的卫生状况，推断食品中肠道 致病菌污染的可能性。
摇试管或换用1支1 mL无菌吸管或吸头反
复吹打，使其混合均匀，制成1:100的样
CHI-SQUARETEST卡方检验详解
CHI-SQUARE TESTAdapted by Anne F. Maben from "Statistics for the Social Sciences" by Vicki SharpThe chi-square (I) test is used to determine whether there is a significant difference between the expected frequencies and the observed frequencies in one or more categories. Do the number of individuals or objects that fall in each category differ significantly from the number you would expect? Is this difference between the expected and observed due to sampling error, or is it a real difference?Chi-Square Test Requirements1. Quantitative data.2. One or more categories.3. Independent observations.4.Adequate sample size (at least 10).5. Simple random sample.6. Data in frequency form.7. All observations must be used.Expected FrequenciesWhen you find the value for chi square, you determine whether the observed frequencies differ significantly from the expected frequencies. You find the expected frequencies for chi square in three ways:I . You hypothesize that all the frequencies are equal in each category. For example, you might expect thathalf of the entering freshmen class of 200 at Tech College will be identified as women and half as men. You figure the expected frequency by dividing the number in the sample by the number of categories. In this exam pie, where there are 200 entering freshmen and two categories, male and female, you divide your sample of 200 by 2, the number of categories, to get 100 (expected frequencies) in each category.2. You determine the expected frequencies on the basis of some prior knowledge. Let's use the Tech Collegeexample again, but this time pretend we have prior knowledge of the frequencies of men and women in each category from last year's entering class, when 60% of the freshmen were men and 40% were women. This year you might expect that 60% of the total would be men and 40% would be women. You find the expected frequencies by multiplying the sample size by each of the hypothesized population proportions. If thefreshmen total were 200, you would expect 120 to be men (60% x 200) and 80 to be women (40% x 200).Now let's take a situation, find the expected frequencies, and use the chi-square test to solve the problem.SituationThai, the manager of a car dealership, did not want to stock cars that were bought less frequently because of their unpopular color. The five colors that he ordered were red, yellow, green, blue, and white. According to Thai, the expected frequencies or number of customers choosing each color should follow the percentages of last year. She felt 20% would choose yellow, 30% would choose red, 10% would choose green, 10% would choose blue, and 30% would choose white. She now took a random sample of 150 customers and asked them their color preferences. The results of this poll are shown in Table 1 under the column labeled observed frequencies."Table 1 - Color Preference for 150 Customers for Thai's Superior Car DealershipCategory Color Observed Frequencies Expected FrequenciesYellow 35 30Red 50 45Green 30 15Blue 10 15White 25 45The expected frequencies in Table 1 are figured from last year's percentages. Based on the percentages for last year, we would expect 20% to choose yellow. Figure the expected frequencies for yellow by taking 20% of the 150 customers, getting an expected frequency of 30 people for this category. For the color red we would expect 30% out of 150 or 45 people to fall in this category. Using this method, Thai figured out the expected frequencies 30, 45, 15, 15, and 45. Obviously, there are discrepancies between the colors preferred by customers in the poll taken by Thai and the colors preferred by the customers who bought their cars last year. Most striking is the difference in the green and white colors. If Thai were to follow the results of her poll, she would stock twice as many green cars than if she were to follow the customer color preference for green based on last year's sales. In the case of white cars, she would stock half as many this year. What to do Thai needs to know whether or not the discrepancies between last year's choices (expected frequencies) and this year's preferences on the basis of his poll (observed frequencies) demonstrate a real change in customer color preferences. It could be that the differences are simply a result of the random sample she chanced to select. If so, then the population of cus-tomers really has not changed from last year as far as color preferences go. The null hypothesis states that there is no significant difference between the expected and observed frequencies. The alternative hypothesis states they are different. The level of significance (the point at which you can say with 95% confidence that the difference is NOT due to chance alone) is set at .05 (the standard for most science experiments.) The chi-square formula used on these data isX2 = (O - E)2where O is the Observed Frequency in each categoryE E is the Expected Frequency in the corresponding categoryis sum ofdf is the "degree of freedom" (n-1)X2 is Chi SquarePROCEDUREWe are now ready to use our formula for X2 and find out if there is a significant difference between the observed and expected frequencies for the customers in choosing cars. We will set up a worksheet; then you will follow the directions to form the columns and solve the formula.1. Directions for Setting Up Worksheet for Chi SquareCategory O E(O - E)(O - E)2(O - E)2Eyellow 35 30 5 25 0.83red 50 45 5 25 0.56green 30 15 15 225 15blue 10 15 -5 25 1.67white 25 45 -20 400 8.89X2 = 26.952. After calculating the Chi Square value, find the "Degrees of Freedom." (DO NOT SQUARE THE NUMBERYOU GET, NOR FIND THE SQUARE ROOT - THE NUMBER YOU GET FROM COMPLETING THECALCULATIONS AS ABOVE IS CHI SQUARE.)Degrees of freedom (df) refers to the number of values that are free to vary after restriction has been placed on the data. For instance, if you have four numbers with the restriction that their sum has to be 50, then three of these numbers can be anything, they are free to vary, but the fourth number definitely isrestricted. For example, the first three numbers could be 15, 20, and 5, adding up to 40; then the fourth number has to be 10 in order that they sum to 50. The degrees of freedom for these values are then three.The degrees of freedom here is defined as N - 1, the number in the group minus one restriction (4 - I ).3. Find the table value for Chi Square. Begin by finding the df found in step 2 along the left hand side of thetable. Run your fingers across the proper row until you reach the predetermined level of significance (.05) atthe column heading on the top of the table. The table value for Chi Square in the correct box of 4 df and P=.05 level of significance is 9.49.4. If the calculated chi-square value for the set of data you are analyzing (26.95) is equal to or greater than thetable value (9.49 ), reject the null hypothesis. There IS a significant difference between the data sets that cannot be due to chance alone. If the number you calculate is LESS than the number you find on the table, than you can probably say that any differences are due to chance alone.In this situation, the rejection of the null hypothesis means that the differences between the expected frequencies (based upon last year's car sales) and the observed frequencies (based upon this year's poll taken by Thai) are not due to chance. That is, they are not due to chance variation in the sample Thai took;there is a real difference between them. Therefore, in deciding what color autos to stock, it would be to Thai's advantage to pay careful attention to the results of her poll!The steps in using the chi-square test may be summarized as follows:Chi-Square I. Write the observed frequencies in column OTest Summary 2. Figure the expected frequencies and write them in column E.3. Use the formula to find the chi-square value:4. Find the df. (N-1)5. Find the table value (consult the Chi Square Table.)6. If your chi-square value is equal to or greater than the table value, reject the nullhypothesis: differences in your data are not due to chance aloneFor example, the reason observed frequencies in a fruit fly genetic breeding lab did not match expected frequencies could be due to such influences as:•Mate selection (certain flies may prefer certain mates)•Too small of a sample size was used•Incorrect identification of male or female flies•The wrong genetic cross was sent from the lab•The flies were mixed in the bottle (carrying unexpected alleles)。
细菌耐药的监测方法
细菌耐药的监测方法一、细菌耐药监测的方法定义 AST 是一个检测细菌耐药性的是一个检测细菌耐药性的体外抑菌试验（ART）AST目的：检出细菌对抗生素的耐药性，预测临床治疗结果。
AST:（1）手工试验 1.纸片扩散法(S,I,R)2.稀释法(MIC)3. E test(MIC)（2）自动仪器 Vitek，Microscan，Phoenix（3）分子试验 PCR直接检测mecA基因（4）酶试验 Nitrocefin、ESBL检测(一)常规药敏试验1.细菌耐药表型检测：判断细菌对抗菌药物的耐药性可根据NCCLS标准，通过测量纸片扩散法、肉汤稀释法和E试验的抑菌圈直径、MIC值和IC值获得。
也可通过以下方法进行检测：（1）耐药筛选试验：以单一药物的单一浓度检测细菌的耐药性被称为耐药筛选试验，临床上常用于筛选耐甲氧西林葡萄球菌、万古霉素中介的葡萄球菌、耐万古霉素肠球菌及氨基糖苷类高水平耐药的肠球菌等。
（2）折点敏感试验：仅用特定的抗菌药物浓度（敏感、中介或耐药折点MIC），而不使用测定MIC时所用的系列对倍稀释抗生素浓度测试细菌对抗菌药物的敏感性，称为折点敏感试验。
（3）双纸片协同试验：双纸片协同试验是主要用于筛选产超广谱β-内酰胺酶（ESBLs）革兰阴性杆菌的纸片琼脂扩散试验。
若指示药敏纸片在朝向阿莫西林/克拉维酸方向有抑菌圈扩大现象（协同），说明测试菌产生超广谱β-内酰胺酶（4）药敏试验的仪器化和自动化：全自动细菌鉴定及药敏分析仪如：Vitek-2、BD-Pheonix、Microscan等运用折点敏感试验的原理可半定量测定抗菌药物的MIC值。
2．β-内酰胺酶检测：主要有碘淀粉测定法（iodometric test）和头孢硝噻吩纸片法（nitrocefin test）。
临床常用头孢硝噻吩纸片法，β-内酰胺酶试验可快速检测流感嗜血杆菌、淋病奈瑟菌、卡他莫拉菌和肠球菌对青霉素的耐药性。
如β-内酰胺酶阳性，表示上述细菌对青霉素、氨苄西林、阿莫西林耐药；表示葡萄球菌和肠球菌对青霉素（包括氨基、羧基和脲基青霉素）耐药。
API20E肠道菌鉴定系统
API 20 E 肠道菌鉴定系统（Ref. 20 100 )布丘菌属：乡间布丘菌Buttiauxella agrestis布戴约维采菌属：水生布戴维约采菌Budvicia aquatica布鲁菌属：布鲁菌属某些种Brucella spp西地西菌属：戴氏西地西菌Cedecea davisae拉氏西地西菌Cedecea lapagei奈氏西地西菌Cedecea neteri柠檬酸杆菌属：无丙二酸柠檬酸杆菌Citrobacter amalonaticus（+/-无丙二酸柠檬酸杆菌）(+/-Citrobacter amalonaticus) 布氏柠檬酸杆菌Citrobacter braakii（+/-弗氏柠檬酸杆菌）(+/-Citrobacter freundii)法氏柠檬酸杆菌Citrobacter farmeri（+/-无丙二酸柠檬酸杆菌）(+/-Citrobacter amalonaticus) 弗氏柠檬酸杆菌Citrobacter freundii（+/-弗氏柠檬酸杆菌）(+/-Citrobacter freundii)柯氏柠檬酸杆菌Citrobacter koseri（=差异柠檬酸杆菌）(=Citrobacter diversus)杨氏柠檬酸杆菌Citrobacter youngae（+/-弗氏柠檬酸杆菌）(+/-Citrobacter freundii)克氏/无丙二酸柠檬酸杆菌Citrobacter koseri/amalonaticus 爱德华菌属：迟钝爱德华菌Edwardsiella tarda保科爱德华菌Edwardsiella hoshinae艾肯菌属：啮蚀艾肯菌Eikenella corrodens肠杆菌属：阴沟肠杆菌Enterobacter cloacae产气肠杆菌Enterobacter aerogenes河生肠杆菌Enterobacter amnigenus中间肠杆菌Enterobacter intermedius日沟维肠杆菌Enterobacter gergoviae阪崎肠杆菌Enterobacter sakazakii生癌肠杆菌Enterobacter cancerogenus（=泰勒肠杆菌）(=Enterobacter taylorae)阿氏肠杆菌Enterobacter asburiae伯克霍尔德菌属：洋葱伯克霍尔德菌Burkholderia cepacie（=洋葱假单胞菌）(=Pseudomonas cepacie)欧文菌属某些种Erwinia spp埃希菌属：大肠埃希菌Escherichia coli弗格森埃希菌Escherichia fergusonii赫氏埃希菌Escherichia hermannii伤口埃希菌Escherichia vulneris假单胞菌属：铜绿假单胞菌Pseudomonsa aeruginosa荧光假单胞菌Pseudomonsa fluorescens恶臭假单胞菌Pseudomonsa putida类鼻疽假单胞菌Pseudomonsa pseudomallei假单胞菌属某些种Pseudomonsa spp寡养单胞菌属：嗜麦寡养食单胞菌Stenotrophomonas maltophilia（=嗜麦芽黄单胞菌）(= Xanthomonas maltophilia)气单胞菌属：嗜水气单胞菌Aeromonas hydrophila豚鼠气单胞菌Aeromonas caviae杀鲑杀鲑气单胞菌Aeromona ssalmonicidas salmonicidas （=杀鲑气单胞菌）(=Aeromonas salmonicidas)温和气单胞菌Aeromonas sobria产碱菌属：产碱菌属某些种Alcaligenes spp克雷伯菌属：植生肺炎克雷伯菌Klebsiella planticola土生拉乌尔菌Raoultella terrigena解鸟氨酸拉乌尔菌Raoultella ornithinolytica产酸克雷伯菌Klebsiella oxytoca肺炎肺炎克雷伯菌Klebsiella pneumoniae pneumoniae臭鼻肺炎克雷伯菌Klebsiella pneumoniae ozaenae鼻硬结肺炎克雷伯菌Klebsiella pneumonia rhinoscleromatis 克吕沃尔菌属：克吕沃尔菌属某些种Kluyvera spp抗坏血酸克吕沃尔菌Kluyvera ascorbata栖冷克吕沃尔菌Kluyvera cryocrescens科泽菌属：特氏科泽菌Koserella trabulsii勒克菌属：非脱羧勒克菌Leclercia adecarboxylata米勒菌属：威斯康星米勒菌Moellerella wisconsensis摩根菌属：摩氏摩根菌Morganella morganii奇异变形杆菌Proteus mirabilis彭氏变形杆菌Proteus penneri普通变形杆菌群Proteus vulgaris group普罗威登斯菌属：产碱普罗威登斯菌Providencia alcalifaciens 雷氏普罗威登斯菌Providencia rettgeri斯氏普罗威登斯菌Providencia stuartii拉氏普罗威登斯菌Providencia rustigianii腐败希瓦菌群Shewanella putrefaciens group 爱文菌属：美洲爱文菌Ewingella americana哈夫尼菌属：蜂房哈夫尼菌Hafnia alvei沙雷菌属：无花果沙雷菌Serratia ficaria居泉沙雷菌Serratia fonticola液化沙雷菌Serratia liquefaciens粘质沙雷菌Serratia marcescens气味沙雷菌1型Serratia odorifera 1气味沙雷菌2型Serratia odorifera 2普城沙雷菌Serratia plymuthica深红沙雷菌Serratia rubidaea志贺菌属：鲍氏志贺菌Shigella bogdii痢疾志贺菌Shigella dysenteriae弗氏志贺菌Shigella flexneri索氏志贺菌Shigella sonnei塔特姆菌属：痰塔特姆菌Tatumella ptyseos志贺菌属某些种Shigella spp耶尔森菌属：小肠结肠炎耶尔森菌Yersinia enterocolitica弗氏耶尔森菌Yersinia fredericksenii中间耶尔森菌Yersinia intermedia克氏耶尔森菌Yersinia kristensenii鼠疫耶尔森菌Yersinia pestis假结核耶尔森菌Yersinia pseudotuberculosis 鲁氏耶尔森菌Yersinia ruckeri莫拉菌属：莫拉菌属某些种Moraxella spp巴斯德菌属：产气巴斯德菌Pasteurella aerogenes溶血曼海姆菌Mannheimia haemolytica多杀巴斯德菌Pasteurella multocida侵肺巴斯德菌Pasteurella pneumotropica巴斯德菌属某些种Pasteurella spp沙门菌属：猪霍乱沙门菌亚利桑那亚种Salmonella choleraesuis ssp arizonae 猪霍乱沙门菌猪霍乱亚种Salmonella choleraesuis ssp choleraesuis 沙门菌甲型副伤寒血清型Salmonella ser.Paratyphi A乙型副伤寒沙门菌Salmonella paratyphi B沙门菌属某些种Salmonella spp伤寒沙门菌Salmonella typhi肠炎沙门菌Salmonella enteritidis沙门菌鸡血清型Salmonella ser.Gallinarum沙门菌鸡白痢血清型Salmonella ser.Pullorum鼠伤寒沙门菌Salmonella typhimurium不动杆菌属：鲍曼不动杆菌Acinetobacter baumannii醋酸钙不动杆菌Acinetobacter calcoaceticus不动杆菌属某些种Acinetobacter spp鞘氨醇单胞菌属：少动鞘氨醇单胞菌Sphlingomonas paucimobilis发光杆菌属：美人鱼发光杆菌Photobacterium damselae（=美人鱼利斯顿菌）(= Listonella damsela)邻单胞菌属：类志贺邻单胞菌Plesiomonas shigelloides拉恩菌属：水生拉恩菌Rahnella aquatilis弧菌属：解藻朊酸弧菌Vibrio alginolyticus霍乱弧菌Vibrio cholerae河流弧菌Vibrio fluvialis霍利斯格里蒙菌Grimontia hollisae最小弧菌Vibrio mimicus副溶血弧菌Vibrio parahaemolyticus创伤弧菌Vibrio vulnificus无色杆菌属：无色杆菌属某些种Achromobacter spp博德特菌属：博德特菌属某些种Bordetella spp色杆菌属：紫色色杆菌Chromobacterium violaceum金色单胞菌属：浅黄假单胞菌Pseudomonas luteola金黄杆菌属：产吲哚金黄杆菌Chryseobacterium indologenes（=产吲哚黄杆菌）(= Flavobacterium indologenes)脑膜脓毒性金黄杆菌Chryseobacterium meningosepticum（=脑膜脓毒性黄杆菌）(=Flavobacterium meningosepticum)黄色单胞菌属：栖稻黄色单胞菌Flavimonas oryzihabitans希瓦菌属：腐败希瓦菌Shewanella putrefaciens鞘氨醇杆菌属：多食鞘氨醇杆菌Sphingobacterium multivorum威克斯菌属：有毒威克斯菌Weeksella virosa动物溃疡威克斯菌Weeksella zoohelcum栖稻假单胞菌Pseudomonas oryzihabitansAPI 20 NE 非肠道菌鉴定系统（Ref. 20 050）假单胞菌属：绿脓假单胞菌Pseudomonas aeruginosa产碱假单胞菌Pseudomonas alcaligenes荧光假单胞菌Pseudomonas fluorescens门多萨假单胞菌Pseudomonas mendocina假产碱假单胞菌Pseudomonas pseudoalcaligenes恶臭假单胞菌Pseudomonas sputita施氏假单胞菌Pseudomonas stutzeri不动杆菌属：鲍氏不动杆菌Acinetobacter baumanii乙酸钙不动杆菌Acinetobacter calcoaceticu溶血不动杆菌Acinetobacter haemolyticus琼氏不动杆菌Acinetobacter junii约氏不动杆菌Acinetobacter johnsonii鲁氏不动杆菌Acinetobacter lwoffii抗辐射不动杆菌Acinetobacter radioresistgns莫拉菌属：腔隙莫拉菌Moraxella lacunata非液化莫拉菌Moraxella nonliquefaciens奥斯陆莫拉菌Moraxella osloensis莫拉菌属某些种Moraxella spp巴斯德菌属：产气巴斯德菌Pasteurella aerogenes溶血曼海姆菌/海藻巴斯德菌Mannheimnia haemolytica/Pasteurella trehalosi 多杀巴斯德菌Pasteurella multocida侵肺巴斯德菌Pasteurella pneumotropica巴斯德菌群EF4 Pasteurella gr.EF4巴斯德菌属某些种Pasteurella spp气单胞菌属：嗜水气单胞菌Aeromonas hydrophila豚鼠气单胞菌Aeromonas caviae温和气单胞菌Aeromonas sobria杀日本鲑杀鲑气单胞菌Aeromonas salmonicida masoucida无色杀鲑气单胞菌Aeromonas salmonicida achromogenes杀鲑杀鲑气单胞菌Aeromonas salmonicida salmonicida（=杀鲑气单胞菌）(= Aeromonsa salmonicida)发光杆菌属：美人鱼发光杆菌Photobacterium damselae（=美人鱼利斯顿菌）( =Listonella damsela)邻单胞菌属：类志贺邻单胞菌Plesiomonas shigelloides威克斯菌属：有毒威克斯菌Weeksella virosa伯格菌属：动物溃疡伯格菌Bergeyella zoohelcum（=动物溃疡威克斯菌）(= Weeksella zoohelcum)窄食单胞菌属：嗜麦芽窄食单胞菌Stenotrophomonas maltophilia（=嗜麦芽黄单胞菌）(=Xanthomonas naltophilia)短波单胞菌属：缺陷短波单胞菌Brevendimonas diminuta（=缺陷假单胞菌）(=Pseudomanas diminuta)泡囊短波单胞菌Brevendimonas vesicularis（=泡囊假单胞菌）(=Pseudomanas vesicularis)腐败希瓦菌群Shewanella putrefaciens group产碱菌属：粪产碱菌Alcaligenes faecalis反硝化无色杆菌Achromobacter denitrificans（=木糖氧化产碱杆菌反硝化亚种）(=A. xylosoxidans subsp denitrificans) 木糖氧化无色杆菌Achromobacter xylosoxidans（=木糖氧化产碱杆菌木糖亚种）(=A. xylosoxidans subsp xylosoxidans) 土壤杆菌属：放射根瘤菌Rhizobium radiobacter博德特菌属：支气管炎博德特菌Bordetella bronchiseptica鸟博德特菌Bordetella aviumCDC：少见沃特菌Wautersia paucula丛毛单胞菌属：睾丸酮丛毛单胞菌Comonas testosteroni（=睾丸酮假单胞菌）(= Pseudomonas testosteroni)食酸代尔夫特菌Delftia acidovorans（=食酸假单胞菌）(= Pseudomonas acidovorans)色杆菌属：紫色色杆菌Chromobacterium violaceum金色单胞菌属：浅黄假单胞菌Pseudomonas luteola黄色单胞菌属：栖稻假单胞菌Pseudomonas oryzihabitans苍白杆菌属：人苍白杆菌Ochrobactrum anthropi寡源杆菌属：解脲寡源杆菌Oligella ureolytica尿道寡源杆菌Oligella urethralis希瓦菌属：腐败希瓦菌Shewanella putrefaciens鞘氨醇单胞菌属：少动鞘氨醇单胞菌Sphlingomonas paucimobilis鞘氨醇杆菌属：多食鞘氨醇杆菌Sphingobacterium multivorum食神鞘氨醇杆菌Sphingobacterium spiritovorum弧菌属：解藻朊酸弧菌Vibrio alginolyticus霍乱弧菌Vibrio cholerae河流弧菌Vibrio fluvialis霍利斯格里蒙菌Grimontia hollisae梅氏弧菌Vibrio metschnikovii最小弧菌Vibrio mimicus副溶血弧菌Vibrio parahaemolyticus创伤弧菌Vibrio vulnificus伯克霍尔德菌属：洋葱伯克霍尔德菌Burkholderia cepacia（=洋葱假单胞菌）(=Pseudomonas cepacia)唐菖蒲伯克霍尔德菌Burkholderia gladioli类鼻疽伯克霍尔德菌Burkholderia pseudomallei（=类鼻疽假单胞菌）(=Pseudomonas pseudomallei)API 20 STREP 链球菌及有关种类的鉴定系统（Ref. 20 600）链球菌属：毗邻颗粒链菌Granulicatella adiacens（=毗邻链球菌）(=Streptococcus adjacens)软弱克养菌Abiotrophia defectiva（=软弱链球菌）(=Streptococcus defectiva)少酸链球菌Streptococcus acidominimus无乳链球菌Streptococcus agalactiae咽峡炎链球菌Streptococcus anginosus牛链球菌ⅠStreptococcus bovis Ⅰ牛链球菌ⅡStreptococcus bovis Ⅱ狗链球菌Streptococcus canis软弱链球菌Streptococcus defectivus停乳停乳链球菌Streptococcus dysgalatiae dysgalatiae （=停乳链球菌）(=Streptococcus dysgalatiae)马肠链球菌Streptococcus equinus似马停乳链球菌Streptococcus dysgalactiae equisimilis (=似马链球菌）(= Streptococcus equisimilis)星座链球菌Streptococcus constellatus中间链球菌Streptococcus intermadius缓症链球菌Streptococcus mitis口腔链球菌Streptococcus oralis类血链球菌Streptococcus parasanguis肺炎链球菌Streptococcus penumoniae豕链球菌Streptococcus porcinus化脓链球菌Streptococcus pyogenes唾液唾液链球菌Streptococcus salivarius salivarius嗜热唾液链球菌Streptococcus salivarius thermophilus 血链球菌Streptococcus sanguinis猪链球菌ⅠStreptococcus suisⅠ猪链球菌ⅡStreptococcus suisⅡ兽瘟马链球菌Streptococcus equi zooepidemicus唾液链球菌Streptococcus salivarius马马链球菌Streptococcus equi equi格氏链球菌Streptococcus gordoniiL群链球菌Streptococcus gr L变异链球菌Streptococcus mutans乳房链球菌Streptococcus uberis肠球菌属：铅黄肠球菌Enterococcus casselifavus耐久肠球菌Enterococcus durans粪肠球菌Enterococcus faecalis屎肠球菌Enterococcus faecium鹑鸡肠球菌Enterococcus gallinarum鸟肠球菌Enterococcus avium气球菌属：浅绿气球菌Aerococcus viridans加德纳菌属：阴道加德纳菌Gardnerella vaginalis孪生球菌属：溶血孪生球菌Gemella haemolysans麻疹孪生球菌Gemella morbillorum乳球菌属：乳脂乳酸乳球菌Lactococcus lactis cremoris乳乳酸乳球菌Lactococcus lactis lactis明串珠菌属：明串珠菌属某些种Luconostoc spp利斯特菌属：无害利斯特菌Listeria innocua伊氏利斯特菌Listeria ivanovii单核细胞增生利斯特菌Listeria monocytogenes斯氏利斯特菌Listeria seeligeri利斯特菌属某些种Listeria spp威氏利斯特菌Listeria welshimeri嗜热链球菌Streptococcus thermophilusAPI CAMPY 弯曲杆菌鉴定系统（Ref. 20 800）弯曲杆菌属：大肠弯曲杆菌Campylobacter coli嗜低温弓形杆菌Arcobacter cryaerohoilus（=嗜低温弯曲杆菌）(= Campylobacter ryaerophilus)胚胎胚胎弯曲杆菌Campylobacter fetus fetus性病胚胎弯曲杆菌Campylobacter fetus venerealis豚肠弯曲杆菌Campylobacter hyointestinalis德莱空肠弯曲杆菌Campylobacter jejuni doylei空肠空肠弯曲杆菌Campylobacter jejuni jejuni红嘴鸥弯曲杆菌Campylobacter lari红嘴鸥弯曲杆菌UPTC Campylobacter lari UPTC粘膜弯曲杆菌Campylobacter mucosalis牛唾液弯曲杆菌Campylobacter sputorrum bubulus唾液弯曲杆菌粪生物变种Campylobacter sputorum bv fecalis 乌普萨拉弯曲杆菌Campylobacter upsaliensis同性恋螺杆菌Helicobacter cinaedi（=同性恋弯曲杆菌）(=Campylobacter cinaedi)芬纳尔螺杆菌Helicobacter fennelliae（=芬纳尔弯曲杆菌）(=Campylobacter fennelliae)幽门螺杆菌Helicobacter pyloriAPI STAPH 葡萄球和微球菌鉴定系统（Ref. 20 500）葡萄球菌属：金黄色葡萄球菌Staphylococcus aureus耳葡萄球菌Staphylococcus auricularis头状葡萄球菌Staphylococcus capitis山羊葡萄球菌Staphylococcus caprae肉葡萄球菌Staphylococcus carnosus产色葡萄球菌Staphylococcus chromogenes科氏科氏葡萄球菌Staphylococcus cohnii cohnii（+/-科氏葡萄球菌）(+/-Staphylococcus cohnii )解脲科氏葡萄球菌Staphylococcus cohnii urealyticum （+/-科氏葡萄球菌）(+/-Staphylococcus cohnii )表皮葡萄球菌Staphylococcus epidermidis溶血葡萄球菌Staphylococcus haemolyticus人葡萄球菌Staphylococcus hominis猪葡萄球菌Staphylococcus hyicus缓慢葡萄球菌Staphylococcus lentus路邓葡萄球菌Staphylococcus lugdunensis腐生葡萄球菌Staphylococcus sarophyticus施氏葡萄球菌Staphylococcus schleiferi松鼠葡萄球菌Staphylococcus sciuri模仿葡萄球菌Staphylococcus simulans沃氏葡萄球菌Staphylococcus warneri木糖葡萄球菌Staphylococcus xylosus库克菌属：克氏库克菌Kocuria kristinae（=克氏微球菌）(=Micrococcus kristinae)变异库克菌Kocuria varians（=变异微球菌）(=Micrococcus varians)玫瑰色库克菌Kocuria roseus（=玫瑰色微球菌）(=Micrococcus roseus)微球菌属：滕黄微球菌Micrococcus luteus莱拉微球菌Micrococcus lylae微球菌属某些种Micrococcus spp粘滑罗斯菌Rothia mucilaginosa不动盖球菌Kytococcus sedentaruis（=不动微球菌）(=Micrococcus sedentaruis)西宫皮肤球菌Dermacoccus nishinomiyaensis（=西宫微球菌）(=Micrococcus nishinomiyaensis)API NH 奈瑟氏菌及嗜血杆菌鉴定系统（Ref. 10 400）伴放线菌素嗜血菌Haemophilus actinomycemtecomitans 奈瑟球菌属：淋病奈瑟球菌Neisseria gonorrhoeae脑膜炎奈瑟球菌Neisseria meningitidis乳糖奈瑟球菌Neisseria lactamica多糖奈瑟球菌Neisseria polysaccharea灰色奈瑟球菌Neisseria cinerea干燥奈瑟球菌Neisseria sicca粘液奈瑟球菌Neisseria mucosa奈瑟菌属某些种Neisseria spp微黄奈瑟球菌Neisseria subflava嗜血杆菌属：流感嗜血杆菌Haemophilus influenzae流感嗜血杆菌ⅠHaemophilus influenzaeⅠ流感嗜血杆菌ⅡHaemophilus influenzaeⅡ流感嗜血杆菌ⅢHaemophilus influenzaeⅢ流感嗜血杆菌ⅣHaemophilus influenzaeⅣ流感嗜血杆菌ⅤHaemophilus influenzaeⅤ副鸡嗜血菌Haemophilus paragallinarum流感嗜血杆菌ⅥHaemophilus influenzaeⅥ流感嗜血杆菌ⅦHaemophilus influenzaeⅦ流感嗜血杆菌ⅧHaemophilus influenzaeⅧ副流感嗜血杆菌Haemophilus parainfluenzae副流感嗜血杆菌ⅠHaemophilus parainfluenzaeⅠ副流感嗜血杆菌ⅡHaemophilus parainfluenzaeⅡ副流感嗜血杆菌ⅢHaemophilus parainfluenzaeⅢ副流感嗜血杆菌ⅣHaemophilus parainfluenzaeⅣ副流感嗜血杆菌ⅤHaemophilus parainfluenzaeⅤ副流感嗜血杆菌ⅥHaemophilus parainfluenzaeⅥ副流感嗜血杆菌ⅦHaemophilus parainfluenzaeⅦ副流感嗜血杆菌ⅧHaemophilus parainfluenzaeⅧ嗜沫嗜血杆菌Haemophilus aphrophilus副嗜沫嗜血杆菌Haemophilus paraphrophilus睡眠嗜组织菌Histophilus somni布兰汉球菌属：卡他莫拉菌（布兰汉菌）Moraxella(Branhamella) catarrhalis 大叶性肺炎放线杆菌Actinobacillus pleuropneumoniaeAPI 20 C AUX 酵母菌鉴定系统（Ref. 20 210）假丝酵母属：白假丝酵母Candida albicans博伊丁假丝酵母Candida boidinii西弗射盾子囊霉Stephanoascus ciferrii软假丝酵母Candida colliculosa都柏林假丝酵母Candida dubliniensis无名假丝酵母Candida famata光滑假丝酵母Candida glabrata季也蒙假丝酵母Candida guilliermondii平常假丝酵母Candida inconspicua乳酒假丝酵母Candida kefyr克柔假丝酵母Candida krusei朗比可假丝酵母Candida lambica解脂假丝酵母Candida lipolytica葡萄牙假丝酵母Candida lusitaniae木篮假丝酵母Candida magnoliae挪威假丝酵母Candida norvegensis近平滑假丝酵母Candida parapsilosis菌膜假丝酵母Candida pelliculosa皱落假丝酵母Candida rugosa圆球形假丝酵母Candida sphaerica热带假丝酵母Candida tropicalis产朊假丝酵母Candida utilis诞沫假丝酵母Candida zeylanoides隐球菌属：浅白隐球酵母Cryptococcus albidus腐殖隐球菌Cryptococcus humicola（=土生假丝酵母）(= Candida humicolus)罗伦隐球酵母Cryptococcus laurentii新型隐球酵母Cryptococcus neoformans地生隐球酵母Cryptococcus terreus指甲隐球酵母Cryptococcus uniguttulatus地霉属：白地霉Geotrichum candidum头状地霉Geotrichum capitatum（=头状丝孢酵母）(=Trichosporon capitatum)克氏地霉Geotrichum klebahnii安格斯毕赤酵母Pichia angusta克勒克酵母属：埃皮斯克勒克酵母Kloeckera apis细尖克勒克酵母Kloeckera apiculata日本克勒克酵母Kloeckera japonica克勒克酵母某些种Kloeckera spp奥默柯达菌Kodamaea ohmeri魏氏原壁菌Prototheca wickerhamii红酵母属：粘红酵母Rhodotorula glutinis小红酵母Rhodotorula minuta粘质红酵母Rhodotorula mucilaginosa（=深红酵母+果蝇红酵母）（=Rhodotorula rubra + Rhodotorula pilimanae）酵母属：酿酒酵母Saccharomyces cerevisiae掷孢酵母属：赭色掷孢酵母Sporobolomyces salmonicolor丝孢酵母属：阿萨丝孢酵母Trichosporon asahii（+/-皮状丝孢酵母）(+/-Trichosporon cutaneum) 墨汁丝孢酵母Trichosporon inkin（+/-皮状丝孢酵母）(+/- Trichosporon cutaneum) 粘性丝孢酵母Trichosporon mucoides（+/-皮状丝孢酵母）(+/- Trichosporon cutaneum) API Candida 假丝酵母鉴定系统（Ref. 10 500）白色假丝酵母Candida albicans无名丝酵母Candida famata光滑假丝酵母Candida glabrata平常假丝酵母Candida inconspicua乳洒假丝酵母Candida kefyr克假丝酵母Candida krusei葡萄牙假丝酵母Candida lusitaniae挪威假丝酵母Candida norvegensis近平滑假丝酵母Candida parapsilosis热带假丝酵母Candida tropicalis新型隐球酵母Cryytococcus neoformans白地霉Geotrichum candidum头状地霉Geotrichum capitatum(=头状丝孢酵母) (=Trichosporon capitatum)酿洒酵母Saccharomyces cerevisiae丝孢酵母某些种Trichosporon sppAPI 20 A 厌氧菌鉴定系统（Ref. 20 300）放线菌属：衣氏放线菌Actinomyces israelii麦氏放线菌Actinomyces mey eri内氏放线菌Actinomyces naeslundii溶齿放线菌Actinomyces odontolyticus粘放线菌Actinomyces viscosus拟杆菌属：粪拟杆菌Bacteroides caccae多毛拟杆菌Bacteroides capillosus吉氏拟杆菌Bacteroides distasonis埃氏拟杆菌Bacteroides eggerthii脆弱拟杆菌Bacteroides fragilis屎拟杆菌Bacteroides merdae卵形拟杆菌Bacteroides ovatus粪便拟杆菌Bacteroides stercoris多型拟杆菌Bacteroides thetaiotaomicron单形拟杆菌Bacteroides uniformis解脲拟杆菌Bacteroides ureolyticus普通拟杆菌Bacteroides vulgatus双歧杆菌属：青春双歧杆菌Bifidobacterium adolescentis双歧双歧杆菌Bifidobacterium bifidum短双歧杆菌Bifidobacterium breve齿双歧杆菌Bifidobacterium dentium双歧杆菌某些种Bifidobacterium spp梭菌属：巴氏梭菌Clostridium barati拜氏梭菌Clostridium beijerinckii双酶梭菌Clostridium bifermentans肉毒梭菌Clostridium botlinum拜氏/丁酸梭菌Clostridium beijerinckii/butyricus 丁酸梭菌Clostridium butyricus尸毒梭菌Clostridium cadaveris梭状梭菌Clostridium clostridioforme艰难梭菌Clostridium difficile矛形梭菌Clostridium hastiforms溶组织梭菌Clostridium histolyticum无害芽胞梭菌Clostridium innocuum泥渣梭菌Clostridium limosum类腐败梭菌Clostridium paraputrificum产气荚膜梭菌Clostridium perfringens多枝梭菌Clostridium ramosum败毒梭菌Clostridium septicum索氏梭菌Clostridium sordellii生孢梭菌Clostridium sporogenes梭菌属某些种Clostridium spp近端梭菌Clostridium subterminate第三梭菌Clostridium tertium破伤风梭菌Clostridium tetani产气柯林斯菌Collinsella aerofaciens真杆菌属：产气真杆菌Eubacterium aerofaciens迟缓埃格特菌Eggerthelle lenta粘液真杆菌Eubacterium limosum梭杆菌属：死亡梭杆菌Fusobacterium mortiferum坏死梭杆菌Fusobacterium necroporum具核梭杆菌Fusobacterium nucleatum可变梭杆菌Fusobacterium varium孪生球菌属：麻疹孪生球菌Gemella mobillorum乳杆菌属：嗜乳酸杆菌Lactobacillus acidophilus发酵乳杆菌Lactobacillus fermentium詹氏乳杆菌Lactobacillus jensenii消化球菌属：黑色消化球菌Peptococcus niger卟啉单胞菌属：不解糖卟啉单胞菌Porphyromonas asaccharolyticus牙龈卟啉单胞菌Porphyromonas gingivalis普雷沃菌属：二路普雷沃菌Prevotella bivia颊普雷沃菌Prevotella buccae解糖胨普雷沃菌Prevotella disiens中间普雷沃菌Prevotella intermedia产黑色普雷沃菌Prevotella melaninogenica口腔普雷沃菌Prevotella oralis口普雷沃菌(=口拟杆菌) Prevotella oris(=Bacteroides oris) 丙酸杆菌属：疮疱丙酸杆菌Propionibacterium acnes颗粒丙酸杆菌Propionibacterium granulosum贪婪丙酸杆菌Propionibacterium avidum丙酸丙酸杆菌Propionibacterium propionicum消化链球菌属：不解糖嗜胨菌Peptoniphilus asaccharolyticus厌氧消化链球菌Peptostreptococcus anaerobius吲哚消化链球菌Peptostreptococcus indolicus大消化链球菌Peptostreptococcus magnus微小消化链球菌Peptostreptococcus micros普氏消化链球菌Peptostreptococcus prevotii消化链球菌群Peptostreptococcus group葡萄球菌属：解糖葡萄球菌Staphylococcus saccharolylicus链球菌属：星座链球菌Streptococcus constellaus中间链球菌Streptococcus intermedius韦荣球菌属：小韦荣球菌Veillonella parvulaAPI CORYNE 棒状杆菌及有关种类的鉴定系统（Ref. 20 600）棒杆菌属：水生雷弗森菌Leifsonia aquatica拥挤棒杆菌Corynebacterium accolens（=G1群棒杆菌）(=Corynebacterium group G1)非发酵棒杆菌Corynebacterium afermentans（+/-ANF-1群棒杆菌）(+/-Corynebacterium group ANF-1) 无枝菌酸棒杆菌Corynebacterium amycolatum银色棒杆菌Corynebacterium argentoratense耳棒杆菌Corynebacterium auris科伊尔棒杆菌Corynebacterium coyleae膀胱炎棒杆菌Corynebacterium cystitidisbelfanti白喉棒杆菌Corynebacterium diphtheriae belfanti 中间白喉棒杆菌 C. diphtheriae intermedius解葡萄糖苷棒杆菌Corynebacterium glucuronolyticum麦氏棒杆菌Corynebacterium macginleyi（=G1/G2群棒杆菌）(= Corynebacterium group G1/G2) 兔肾棒杆菌群Corynebacterium renale group丙酸棒杆菌Corynebacterium propinquum(=ANF-3群棒杆菌) (= Corynebacterium ANF-3)解葡萄糖苷棒杆菌Corynebacterium glucuronolyticum解脲棒杆菌Corynebacterium urealyticum(=D2群棒杆菌) (= Corynebacterium Group D2)牛棒杆菌Corynebacterium bovis缓和白喉棒杆菌Corynebacterium diphtheriae mitis重白喉棒杆菌Corynebacterium diphtheriae gravis杰氏棒杆菌Corynebacterium jeikeium库氏棒杆菌Corynebacterium kutscher（=JK群棒杆菌）(= Corynebacterium gr.JK)极小棒杆菌Corynebacterium minutissimum多毛棒杆菌Corynebacterium pilosum假结核棒杆菌Corynebacterium pseudotuberculosis假白喉棒杆菌Corynebacterium pseudodiphtheriticum兔肾炎棒杆菌Corynebacterium renale纹带棒杆菌Corynebacterium striatum溃疡棒杆菌Corynebacterium ulceransF-1群棒杆菌Corynebacterium group F- 1（=F群棒杆菌）(= Corynebacterium group F)G群棒杆菌Corynebacterium group G（=G1/G2群棒杆菌）(= Corynebacterium group G1/G2) 红球菌属：马红球菌Rhodococcus equi红球菌属某些种Rhodococcus spp利斯特菌属：单核细胞增生利斯特菌Listeria monocytogenes无害利斯特菌Listeria innocua格氏利斯特菌Listeria grayi斯氏利斯特菌Listeria seeligeri魏氏利斯特菌Listeria welshimeri利斯特菌属某些种Listeria spp厄菌属：厄菌属某些种Oerskovia spp放线菌属：无硝纽氏放线菌Actinomyces neuii anitratus纽氏纽氏放线菌Actinomyces neuii neuiiActinomyces neuii radingae图列茨放线菌Actinomyces neuii turicensis丹毒丝菌属：猪红斑丹毒丝菌Erysipelothrix rhusiopathiae隐秘杆菌属：溶血隐秘杆菌Arcanobacterium haemolyticum伯纳德隐秘杆菌Arcanobacterium bernardiae化脓隐秘杆菌Arcanobacterium pyogenes（=化脓放线菌）(= Actinomyces pyogenes)短杆菌属：短杆菌属某些种Brevibacterium spp乳酪短杆菌Brevibacterium casei（=B1和B3群棒杆菌）(= Corynebacterium gr B1 and B3) 表皮短杆菌Brevibacterium epidermidis（=B群棒杆菌）(= corynebacterium gr B)加德纳菌属：阴道加德纳菌Gardnerella vaginalis戈登菌属：戈登菌属某些种Gordona spp罗菌属：龋齿罗菌Rothia dentocariosa节杆菌属：节杆菌属某些种Arthrobacter spp（=A群棒杆菌）(= Corynebacterium gr.A)纤维单胞菌属：纤维单胞菌属某些种Cellulomonas spp（=A3/A4群棒杆菌）(= Corynebacterium gr A3/A4)特氏纤维单胞菌Cellulomonas turbata（=特氏厄菌）(=Oerskovia turbata)皮肤杆菌属：人皮肤杆菌Dermabacter hominis迪茨菌属：迪茨菌属某些种Dietzia spp微小杆菌属：微小杆菌属某些种Microbacterium spp（+/-A4/A5群棒杆菌）(+/- Corynebacterium gr.A4/A5)奴卡菌属：奴卡菌属某些种Nocardia spp厄菌属：纤维化纤维微细菌Cellulosimicrobium cellulans丙酸杆菌属：痤疮丙酸杆菌Propionibacterium acnes贪婪丙酸杆菌Propionibacterium avidum微杆菌属某些种Microbacterium sppAPI 10 S 肠杆菌科快速筛选鉴定系统（Ref. 10 100 ）鲍曼不动杆菌Acinetobacter baumannii嗜水气单胞菌Aeromonas hydrophilia产吲哚金黄杆菌Chryseobacterium indologenes（=产吲哚黄杆菌）(=Flavobacterium indologenes)脑膜脓毒性金黄杆菌Chryseobacterium meningosepticum （=脑膜脓毒性黄杆菌）(=Flvobacterium meningosepticum) 布氏柠檬酸杆菌Citrobacter braakii（+/-弗氏柠檬酸杆菌）(+/- Citrobacter freundii)法氏柠檬酸杆菌Citrobacter farmeri（+/-无丙二酸柠檬酸杆菌）(+/-Citrobacter amalonaticus)弗氏柠檬酸杆菌Citrobacter freundii（+/-弗氏柠檬酸杆菌）(+/-Citrobacter freundii)克氏/无丙二酸柠檬酸杆菌Citrobacter koseri/amanlonaticus 迟钝爱德华菌Edwardsiella tarda肠杆菌属某些种Enterobacter spp产气肠杆菌Enterobacter aerogenes河生肠杆菌Enterobacter amnigenus阴沟肠杆菌Enterobacter cloacae大肠埃希菌Escherichia coli伤口埃菌Escherichia vulneris蜂房哈夫尼亚菌Hafnia alvei产酸克雷伯菌Klebsiella oxytoca肺炎肺炎克雷伯菌Klebsiella pneumoniae pneumoniae摩根摩根菌Morganella morganii泛菌属某些种Pantoea spp（+/-成团肠杆菌）(+/- Enterobacter agglomerans)类志贺邻单胞菌Plesiomonas shigelloides奇异变形杆菌Proteus mirabilis彭氏变形杆菌Proteus penneri普通变形菌群Proteus vulgaris group普通变形杆菌Proteus vulgaris雷氏普罗威登斯菌Providencia rettgeri斯氏/产碱普罗威登斯菌Providencia stuartii/ alcalifaciens铜绿假单胞菌Pseudomonas aeruginosa荧光假单胞菌Pseudomonas fluorescens恶臭假单胞菌Pseudomonas putida假单胞菌属某些种Pseudomonas spp猪霍乱沙门菌亚利桑那亚种Salmonella choleraesuis ssp arizonae 猪霍乱沙门菌猪霍乱亚种Salmonella choleraesuis ssp choleraesuis 沙门菌鸡血清型Salmonella ser.Gallinarum沙门菌甲型副伤寒血清型Salmonella ser.Paratyphi A沙门菌鸡白痢血清型Salmonella ser.Pullorum沙门菌属某些种Salmonella spp伤寒沙门菌Salmonella typhi液化沙雷菌Serratia liquefaciens粘质沙雷菌Serratia marcescens气味沙雷菌Serratia odorifera腐败希瓦菌Shewanella putrefaciens志贺菌属某些种Shigella spp多食鞘氨醇杆菌Sphingobacterium multivorum嗜麦芽寡养单胞菌Stenotrophomonus maltophilia（=嗜麦芽黄单胞菌）(=Xanthomonas maltophilia)解藻朊酸弧菌Vibrio alginolyticus副溶血弧菌Vibrio parahaemolyticus创伤弧菌Vibrio vulnificus霍乱弧菌Vibrio cholerae小肠结肠类耶尔森菌V ersinia enterocolitica假结核耶尔森菌Versinia pseudotuberculosis腐败希瓦菌群Shewanella putrefaciens groupRAPID 20 E 肠道菌快速鉴定系统（Ref. 20 700）柠檬酸杆菌属：无丙二酸柠檬酸杆菌Citrobacter amalonaticua（+/-无丙二酸柠檬酸杆菌）(+/-Citrobacter amalonaticus)法氏柠檬酸杆菌Citrobacter farmeri（+/-无丙二酸柠檬酸杆菌）(+/- C. Amalonaticus)弗氏柠檬酸杆菌群Citrobacter freundii group（=弗氏柠檬酸杆菌）(=C. Freunclii)克氏柠檬酸杆菌Citrobacter Koseri（=差异柠檬酸杆菌）(= C.diversus)爱德华菌属：保科爱德华菌Edwardsiella hoshinae迟钝爱德华菌Edwardsiella tarda肠杆菌属：产气肠杆菌Enterobacter aerogenes阿氏肠杆菌Enterobacter asburiae生癌肠杆菌Enterobacter cancerogenus（=泰勒肠杆菌）(=Enterobacter taylorae)阴沟肠杆菌Enterobacter cloacae日沟维肠杆菌Enterobacter gergoviae阪崎肠杆菌Enterobacter sakazakii埃希菌属：大肠埃希菌Escherichia coli费格森埃希菌Escherichia fergusonii赫氏埃希菌Escherichia hermannii伤口埃希菌Escherichia vulneris爱文菌属：美洲爱文菌Ewingella americana哈夫尼亚菌属：蜂房哈夫尼菌Hafnia alvei克雷伯菌属：解鸟氨酸拉乌尔菌Raoultella ornithinolytica产酸克雷伯菌Klebsiella oxytoca植生克雷伯菌Klebsiella planticola臭鼻肺炎克雷伯菌Klebsiella pneumoniae ozaenae肺炎肺炎克雷伯菌Klebsiella pneumoniae pneumoniae鼻硬结肺炎克雷伯菌Klebsiella pneumoniae rhinoscieromatis 土生克雷伯菌Klebsiella terrigena克吕沃尔菌属：抗坏血酸克吕沃尔菌Kluyvera ascorbata栖冷克吕沃尔菌Kluyvera cryocrescens克吕沃尔菌属某些种Kluyvera spp米勒菌属：威斯康星米勒菌Moellerella wisconsensis摩根菌属：摩氏摩根菌Morganella morganii勒克菌属：非脱羧勒克菌Leclercia adcarboxglata变形杆菌属：奇异变形菌Proteus mirabilis彭氏变形菌Proteus penneri普通变形菌群Proteus vuigaris group泛菌属：泛菌属某些种Pantoea spp（+/-成团肠杆菌）(+/-Enterobacter agglonerans)发光杆菌属：美人鱼发光杆菌Photobacterium danselae（=美人鱼利斯顿菌）(=Listonella damsela)普罗威登斯菌属：产碱普罗威登斯菌Providencia alcalifaciens雷氏普罗威登斯菌Providencia rettgeri斯氏普罗威登斯菌Providencia stuartii沙门菌属：猪霍乱沙门菌亚利桑那亚种Salmonella choleraesuis ssp arizonae猪霍乱沙门菌猪霍乱亚种Salmonella choleraesuis ssp choleraesuis沙门菌甲型副伤寒血清型Salmonella ser.Paratyphi A沙门属某些种Salmonella spp伤寒沙门菌Salmonella typhi鸡沙门菌Salmonella gallinarum志贺菌属：志贺菌属某些捉Shigella spp索氏志贺菌Shigella sonnei沙雷菌属：无花果沙雷菌Serratia ficaria居泉沙雷菌Serratia fonticola液化沙雷菌Serratia liquefaciens粘质沙雷菌Serratia marcescens气味沙雷菌Serratia odorifera普城沙雷菌Serratia plymuthica深红沙雷菌Serratia rubidaea寡养单胞菌属：嗜麦芽寡养单胞菌Stenotrophomonas maltophilia（=嗜麦芽黄单胞菌）(=Xanthomonas maltophilia)耶尔森菌属：小肠结肠炎耶尔森菌Yersinia enterocolitica鼠疫耶尔森菌Yersinia pestis假结核耶尔森菌Yersinia pseudotuberculosis不动杆菌属/假单胞菌属：不动杆菌属某些种Acinetobacter spp不动杆菌属某些种/假单胞菌属某些种Acinetobacter spp/Pseudomonas spp 气单胞菌属：嗜水气单胞菌Aeromonas hydrophila邻单抱菌属：类志贺邻单胞菌Plesiomonas shigelloides弧菌属：解藻朊酸弧菌Vibrio alginolyticus霍乱弧菌Vibrio cholerae河流孤菌Vibrio fluvialis霍利斯格里蒙菌Grimontia hollisae梅氏弧菌Vibrio metschnikovi副溶血弧菌Vibrio parahaemolyticus创伤弧菌Vibrio vulnificus洋葱伯克霍尔德菌Burkholderia cepacia（=洋葱假单胞菌）(=Pseudomonas cepacia) 乡间布丘菌Buttiauxella agrestis戴氏西地西菌Cedecea davisae拉氏西地西菌Cedecea lapagei奈氏西地西菌Cedecea neteri西地西菌属某些种Cedecea sppAPI LISTERIA 李斯特菌鉴定系统（Ref. 10 300）李斯特菌属：单核细胞增生利斯特菌Listeria monocytogenes 无害利斯特菌Listeria innocua伊氏利斯特菌Listeria ivanovii威氏利斯特菌Listeria welshimeri斯氏利斯特菌Listeria seeligeri格氏利斯特菌Listeria grayi。
大肠埃希菌检查法
大肠埃希菌检查法Escherichia Coli Test Method1.目的建立大肠埃希菌检查法的标准操作程序。
2.围适用于大肠埃希菌检查的操作。
3.责任者QC化验员。
4.程序：•大肠埃希菌(Escherichia coli)即大肠埃希菌，为肠埃希菌科埃希菌属的模式种。
埃希菌属除大肠埃希菌外，新近发现有非脱竣埃希氏菌等5个种。
大肠埃希菌是人和温血动物肠道的栖居菌，随粪便排出体外。
在药品中检出大肠埃希菌，说明该样品受到人和温血动物的粪便污染，即可能污染肠道病原体。
大肠埃希菌除普通大肠埃希菌外尚有致病性大肠埃希菌，可引起婴幼儿、成人爆发性腹泻。
为保证人体安康，口服药品必须检查大肠埃希菌。
・用4-甲基伞形酮葡糖昔酸(4-Methylumbelliferyl- 0 -D-glucuronide, MUG) 和靛基质(Indole)试验检查大肠埃希菌是一项新技术，其检验步骤为：增菌培养后，转种MUG-蛋白淼培养基培养，多数情况下不需要从混合菌中别离单个菌，如MUG、Indole试验为阳性或阴性即可报告结果。
・原理：利用目标菌限定酶作用的底物的水解产物，产生颜色或荧光反响作为指示系统来鉴定目标菌。
实验证明，96%的大肠埃希菌含B-葡糖昔酸酶(3 -glucuronidase, GUD), 约10%的沙门菌属一些菌种也含有此酶。
MUG被GUD水解，产生荧光，由于荧光反响的敏感度较颜色反响强千万倍，易于观察，没有主观性，因而用MUG鉴定大肠埃希菌已被广泛应用于临床、食品.饮水、污水等的检测。
单一的MUG鉴别大肠埃希菌其漏检率达6%,鉴于98%的大肠埃希菌其靛基质试验为阳性，故将MUG与靛基质试验结合，比单用MUG可提高大肠埃希菌的检出率。
如MUG与Indole 试验的反响不一致时，那么需将供试液的增菌培养物用EMB琼脂平板别离培养. 革兰染色、镜检及生化试验鉴别。
该法理论上可使大肠埃希菌的检出率达98%。
如仅用IMViC生化试验来鉴别大肠埃希菌属中的大肠埃希菌，其结果是含混的。
医学检验微生物api板条简介及原理
API 20 E
• 革兰氏阴性杆菌鉴定 • 用於鉴定环境 / 食品样本中的分离菌株 • 肠道杆菌的鉴定经典方法 • 24小时出结果 (Rapid 20E : 4 小时)
API 20 E
1 2 4 1 2 4 1 2 4 1 2 4 1 2 4 1 2 4 1 2 4 1 2 4 12 4
ONPG ADH LDC ODC CIT H2S UREE TDA IND VP GEL GLU MAN INO SOR RHA SAC MEL AMY ARA OX NO2 N2 MOBMcC OF-O OF-F
种能抵抗恶劣生长条件。 • 硫化物还原厌氧菌可作为粪便污染的指标。 • 肉毒梭菌及产菌荚膜梭菌为主要食品致病菌
。 • 需厌氧条件下孵育试条（产气盒/袋，石腊油
）。
API 20 A
厌氧菌
• 利用 21个生化反应鉴 定厌氧菌
• 必须进行额外试验
• 原理
待检菌株经 24/48 小 时厌氧培养后产生的 代谢物由试条里的pH 显示剂检测出结果
• 链球菌普遍存在於人及环境中。 • 粪链球菌作为水质粪便污染的指标。 • 47 个条目
API 20 STREP
链球菌鉴定
利用传统生化及发酵反应 鉴定链球菌
肉眼判读结果
原理
使用20个风乾底物，包括11 个发酵试验及9个传统生化/酶 反应
APILAB
API 20 STREP
4h 1
• Pasteurella
(巴斯德氏菌)
• Chromobacter
(色杆菌)
• Chromobacterium (色杆菌属)
• Pseudomonas
(假单胞菌属)
(3) • Bordetella/Alcaligenes (1)
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Report 检验报告
OUR REF NO: ATS/MED/77/04/pl
DATE: 31 March, 2004 Page 1 of 2
COMPANY: Okada Ecotech Pte Ltd
Blk 1 Pasir Panjang Road
Distripark #07-15/17
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文件编号：ATS/MED/77/04/pl
日期： 2004年3月31日第1/2页
送检公司：冈田生态技术私人有限公司
地址：新加坡亚历山大分销园长滩路1区07－15/17号
邮编： 118478
DATE SAMPLE RECEIVED: 23 March, 2004
DATE ANALYZED: 23 March, 20004
DATE TEST COMPLETED: 31 March, 2004
SAMPLE DESCRIPTION: One bottle said to be Biovectrol 20EM (Indoor) was
received with references :-
2004030801
BN
送检日期： 2004年3月23日
检验日期： 2004年3月23日
完成日期： 2004年3月31日
样品描述：名称为Biovectrol 20EM（室内用）样品一瓶，样品号：
BN
2004030801
Method: Chemical Disinfectants & antiseptics – Basic Bactericidal Activity (BS EN 1040:1997)
检验方法：化学消毒剂和杀菌剂 – 基础试验（BS EN 1040:1997）
Procedure:
检验程序：
1. 1ml of sample was transferred to 79ml of sterile water to make dilution of 1:79.
1. 将1毫升样品掺入79毫升的无菌水中，稀释比例为1:79。
2. 8ml of diluted samples was transferred to 50ml sterile tubes and 1ml of water was added.
2. 将8毫升上述稀释样品倒入50毫升的无菌试管中，再加入1毫升水。
3. 1ml of the bacterial test suspension was inoculated into the tube.
3. 将1毫升的细菌悬液注入无菌试管中。
4. The test mixture was mixed well and placed in 20ºC ± 1ºC water bath.
4. 混合均匀后，将试管置于20ºC ± 1ºC的水槽中。
5. After 40min and 60min ± 10sec contact time, 1ml of the test mixture was transferred into a tube containing 8ml neutralizer and 1ml water.
5. 分别静置40分钟及60分钟 ± 10秒后，将1毫升混合液注入盛有8毫升中和剂和1毫升水的试管中。
6. The test mixture was mixed well and placed in 20ºC ± 1ºC water bath.
6. 混合均匀后，将试管置于20ºC ± 1ºC的水槽中。
7. After a neutralization time of 5mins ± 10sec, 1ml of the mixture was tested immediately using pour-plate method with Tryptone Soya Agar (TSA).
7. 中和时间至5分钟 ± 10秒后，将1毫升混合液取出，立即用胰蛋白胨大豆琼脂(TSA) 通过平皿灌浇法进行试验。
8. The plates were incubated at 35ºC for 48 hours.
8. 将平皿置于35ºC培养48小时。
9. This procedure was repeated for the other bacterial test suspensions.
9. 此程序亦重复应用于其它细菌悬液。
Results: On analysis, the following results were obtained:
试验结果：经试验分析得出以下结果：
Test organism 测试细菌类型Initial Bacterial Load (cfu/ml)
最初菌落群数
Staphylococcus aureus (ATCC 6538)
金黄色葡萄球菌
3.80 x 108 Pseudomonas aeruginosa (ATCC 15442)
绿脓杆菌
1.55 x 108 Salmonella typhimurium (ATCC 14028)
鼠伤寒沙门氏菌
3.20 x 108 Escherichia coli (ATCC 25922)
大肠埃希杆菌
1.31 x 108
OUR REF NO: ATS/MED/77/04/pl 31 March, 2004
文件编号： ATS/MED/77/04/pl 2004年3月31日COMPANY: Okada Ecotech Pte Ltd Page 2 of 2
送检公司: 冈田生态技术私人有限公司第2/2页
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TEST RESULTS FOR SAMPLE AT DILUTION OF 1:79
稀释比例为1:79的样品试验结果
Test organism 测试细菌类型After contact time
of 40 mins
静置40分钟后
After contact time of 60 mins
静置60分钟后
Observation of
plates
平皿观察
Total Viable
Count (cfu/ml)
活细菌数合计
Observation of plates
平皿观察
Total Viable
Count (cfu/ml)
活细菌数合计
Staphylococcus aureus 金黄色葡萄球菌Colonies observed
观察到菌落
1.20 x 106
Colonies observed
观察到菌落
1.05 x 106
Pseudomonas aeruginosa 绿脓杆菌Colonies observed
观察到菌落
1.50 x 106
No Colonies observed
未观察到菌落
1.40 x 106
Salmonella typhimurium 鼠伤寒沙门氏菌Colonies observed
观察到菌落
3.60 x 104
Colonies observed
观察到菌落
2.20 x 103
Escherichia coli 大肠埃希杆菌Colonies observed
观察到菌落
80
No Colonies observed
未观察到菌落
30
Test organism 测试细菌类型
Reduction in viability (% killed)
菌落群数减少率(杀死率%)
After contact time of 40 mins
静置40分钟后
After contact time of 60 mins
静置60分钟后
Staphylococcus aureus
金黄色葡萄球菌
99.68% 99.72% Pseudomonas aeruginosa
绿脓杆菌
99.03% 99.10%
Salmonella typhimurium 鼠伤寒沙门氏菌99.99% >
99.99%
Escherichia coli 大肠埃希杆菌>99.99% >
99.99%
NOTES:
注：
cfu – colony forming unit cfu – 菌落形成单位。