2019级检验英语演示文稿7-精选文档22页

To utilize this assay, one capture antibody is purified and bound to a solid phase typically attached to the bottom of a plate well. Antigen is then added and allowed to complex with the bound antibody. Unbound products are then removed with a wash, and a labeled second detection antibody is allowed to bind to the antigen, thus completing the sandwich.
妊娠；受孕 绒毛膜的 促性腺激素 乳汁；胶乳 多聚物 绒毛膜上皮癌 包虫囊 搅拌；动 精力旺盛的；强壮的
Lesson Seven
Pregnancy Testing
PRINCIPLE: Human chorionic gonadotropin (HCG) appears in the urine of pregnant females, and determination of its presence generally indicates a positive test for pregnancy .
About immunology
immune response
免疫反应
antigen---antibody
monoclonal antibody (McAb) 单克隆抗体
polyclonal antibody
多克隆抗体
recombinant antigen
重组抗原
serial dilutions
系列稀释
radioimmunoassay 放射免疫方法 immunoblotting: PAGE 聚丙烯酰胺凝胶电泳
polyacrylamide gel electrophoresis
About molecular biology
immunofluorescence
免疫荧光
polymerase chain reaction (PCR) 聚合酶联反应
Lesson Seven
Pregnancy Testing
REAGENTS: Available as a kit from Roche Diagnostics .
PROCEDURE: 1. Drop 1 drop urine in circle on glass slide . 2. Add 1 drop antiserum reagent . 3. Stir and rotate gently for 30 sec . 4. Add 1 drop antigen reagent . 5. Mix over entire circle . 6. Rotate slide vigorously for no longer than 2 min while observing for agglutination .
the secondary antibody 二抗
the absorbance
吸光度
Lesson Seven
Pregnancy Testing
pregnancy chorionic gonadotropin latex polymer chorioepithelioma hydatid stir vigorously
high titer
高滴度
enzyme-linked immunosorbent assay (ELISA )
酶联免疫吸附试验
horseradish peroxidase-conjugated goat anti-rabbit IgG
辣根过氧化物酶标记的羊抗兔IgG
About immunology
Lesson Seven
Pregnancy Testing
PRINCIPLE: Patients with chorioepithelioma or hydatid mole(葡萄胎) frequently excrete HCG in the urine. Therefore a positive result obtained when testing these patients does not always indicate a positive test for pregnancy.
Lesson Seven
Pregnancy Testing
INTERPRETATION: Report as 1. Positive test: No agglutination within 2 min. HCG present in the urine of pregnant women will inhibit agglutination of latex particles in the antigen reagent, resulting in a smooth, milky appearance after rotation.
Sandwich ELISA
This can be accomplished with either two monoclonal antibodies that recognize discrete (不连续的) sites or one batch of affinity-purified polyclonal antibodies.
Sandwich ELISA
To quantitate Ag-specific IgG, serial dilutions of serum prepared from tail vein blood were added to ELISA plates coated with rSRS9 or rSAG1(50ng/well). After incubation(孵育) with peroxidase-coupled goat anti-mouse IgG and its substrate( 底 物 ), and plates were read colorimetrically at 450 nm.
precipitation : double diffusion test
双扩散试验
gel immunoelectrophoresis
凝胶免疫电泳
agglutination: direct agglutination 直接凝集试验 indirect agglutination间接凝集试验
latex agglutination test; indirect hemagglutination test
restriction fragment-length polymorphism analysis
(RFLP) 限制性片段多态性分析
restriction endonuclease: BamH Ⅰ; EcoRⅠ
限制性内切酶
(5 ´-GAATTC-3´)
Western-blot 免疫印迹
(3 ´-CTTAAG-5´)
Lesson Seven
Pregnancy Testing
PRINCIPLE: Test components consist of an antigen in the form of suspension of HCG chemically bonded to latex polymer particles and an antiserum containing antibodies to HCG.
anti-HIV McAb
ELISA
ELISA
positive conห้องสมุดไป่ตู้rol
阳性对照
negative control
阴性对照
blank control
空白对照
coating overnight at 4C 包被过夜
blocked with
封闭
washed with 0.1% Tween 20 in PBS
Lesson Seven
Pregnancy Testing
PRINCIPLE: Utilizing an immunologic technique based on the principle of indirect agglutination, tests will detect HCG at levels of 1.5 to 2.5 International Units (IU) per milliliter. Since levels as low as 1.5 IU have been reported as early as 5 days after the first missed period, negative tests should be repeated within 1 to 2 weeks.
Lesson Seven
Pregnancy Testing
INTERPRETATION: Report as 2. Negative test: Any degree of agglutination within 2 min. Lack of HCG in the urine of nonpregnant women permits agglutination of the antigen and antiserum reagents within a 2 min period, producing easily recognizable clumping(块) of the latex particles.
Lesson Seven
Pregnancy Testing
PRINCIPLE: When the antiserum is mixed with urine containing a detectable level of HCG, it is neutralized, and no agglutination occurs. This is a positive test for pregnancy. Urine that contains no HCG cannot neutralize the antiserum, resulting in agglutination of the latex reagent. This indicates a negative test for pregnancy.
Sandwich ELISA
One of the most useful of the immunoassays is the two antibody sandwich ELISA. This assay is used to determine the antigen concentration in unknown samples. This ELISA is fast and accurate, and if a purified antigen standard is available, the assay can determine the absolute amount of antigen in an unknown sample. The sandwich ELISA requires two antibodies that bind to epitopes(表位) that do not overlap on the antigen.
Southren-blot (DNA-DNA hybridization)
Northern-blot (DNA-RNA hybridization)
DNA probe DNA探针
Immunochromatography
免疫层析
immunogold
immunogold labelling rHIV-Ag1 rHIV-Ag2
Sandwich ELISA
The assay is then quantitated by measuring the amount of labeled second antibody bound to the matrix (矩阵), through the use of a colorimetric substrate. Major advantages of this technique are that the antigen does not need to be purified prior to use, and that these assays are very specific. However, one disadvantage is that not all antibodies can be used. Monoclonal antibody combinations must be qualified as unattached pairs, meaning that they can recognize separate epitopes on the antigen so they do not hinder each other binding.