尿微量白蛋白(MAU)测定标准操作程序SOP文件
尿液蛋白质测定(手工法)操作规程sop
尿液蛋白质测定（手工法）操作规程sop
一、项目名称：尿液蛋白质定性检查
二、方法：磺基水杨酸法
三、原理：磺基水杨酸为生物碱试剂，在酸性环境下，其阴离子可
与带正电荷的蛋白质结合成不溶性蛋白盐而沉淀。
四、试剂：10%磺基水杨酸乙醇溶液。
五、仪器：手工法
六、操作步骤：
1、取试管加尿液3ml。
2、滴加10%磺基水杨酸溶液3－4滴，形成界面。
3、如尿呈浑浊，表示有蛋白质存在，混浊深浅表示含量的多
少。
七、结果判断：
●阴性：尿液外观仍清晰透明，不呈浑浊。
●微量（+/-）：轻微浑浊，隐约可见。
●阳性（+）：明显的白色混浊，但无颗粒出现。
（++）：稀薄乳样浑浊，出现颗粒。
（+++）：混浊，有絮片状沉淀。
（++++）：絮状混浊，有大凝块下沉。
八、参考范围：正常人为阴性。
九、标本要求：
1、标本应新鲜，并及时送检。
2、尿液标本应避免经血、白带、精液、粪便等混入。
3、容器应清洁、干燥。
十、临床意义：分为功能性、体位性、病理性蛋白尿，后者见于肾
炎、肾病综合症等。
十一、注意事项：
1、此法比较敏感。
2、如尿液浑浊，应先离心或过滤。
3、强碱性尿可出现假阴性，应加5%醋酸溶液数滴酸化后再做试
验。
4、有机碘造影剂、超大剂量使用青霉素等均可致假阳性。
5、尿中含高浓度尿酸或尿酸盐时，可呈假阳性。
十二、参考文献：
全国临床检验操作规程（第3版）
十三、
编写者：
制定日期：
修改日期：
科主任对规程认可：。
尿微量白蛋白(MAU)测定试剂盒(荧光免疫层析法)产品技术要求sz
尿微量白蛋白（MAU）测定试剂盒（荧光免疫层析法）
型号、规格
20人份/盒， 40人份/盒，60人份/盒
结构及组成
试剂盒由检测卡和ID卡组成；检测卡主要组成成分有硝酸纤维素膜，玻璃纤维素膜，吸水纸，PVC板；其中硝酸纤维素膜在特定位置上包被有白蛋白抗原和兔IgG，玻璃纤维素膜上喷有荧光微球标记的白蛋白单克隆抗体和抗兔IgG抗体；ID卡包含校准曲线和批号。
试剂的线性范围为5mg/L～100mg/L，在此线性范围内：线性相关系数r应不小于0.9900。
2.3检测限
检测限应不大于5mg/L。
2.4批内差精密度
变异系数（CV）应≤15%。
2Hale Waihona Puke 5批间差变异系数（CV）应≤15%。
2.6准确度
回收率应在 85%～115%范围内。
1/1
产品适用范围/预期用途
本试剂盒用于体外定量测定人尿液中微量白蛋白（MAU）的含量。临床上主要用于肾脏疾病的辅助诊断。
2.性能指标
2.1物理性状
2.1.1外观
试剂卡外观平整，材料附着牢固。
2.1.2膜条宽度
膜条宽度为4.00±0.40mm。
2.1.3液体移行速度
液体移行速度应不低于25mm/min。
2.2线性范围
尿微量蛋白操作流程
尿微量蛋白操作流程1. 简介尿微量蛋白是一种常用的临床检测方法，用于检测尿液中是否存在微量的蛋白质。
本文将详细描述尿微量蛋白的操作流程，包括样本收集、处理和分析等步骤。
2. 实验材料和设备准备•尿液收集容器•尿液离心管•显微镜•尿液分析试纸•生化分析仪器（如比色计）•实验室耗材（试剂、移液管、离心机等）3. 样本收集1.提醒受试者在收集尿样前先进行外部清洁，以避免外源性污染。
2.提供干净的尿液收集容器，并告知受试者在收集尿样时避免接触容器内壁。
3.受试者应将第一次排尿舍弃，从第二次开始进行采样。
4.受试者应采集早晨第一次排尿作为样本，并将其放入标有姓名和日期的离心管中。
4. 样本处理1.将采集好的尿液样本离心，以去除其中的悬浮物和颗粒。
2.将离心后的尿液转移到一个干净的容器中，避免留下任何固体残留物。
3.对于需要长时间保存的样本，可以加入一定量的保存液（如硼酸盐缓冲液）。
5. 样本分析1.尿微量蛋白分析可以通过尿液分析试纸或生化分析仪器进行。
以下将分别介绍两种方法。
5.1 使用尿液分析试纸1.取出一张尿液分析试纸，并查看其有效期和使用说明。
2.使用移液管或者特定的取样器，在尿样中吸取适量的尿液，并滴在试纸上相应位置。
3.等待指定时间（通常为几十秒至数分钟），使试纸与尿样充分反应。
4.使用试纸上标示的颜色比对图，根据颜色变化确定蛋白质含量。
5.2 使用生化分析仪器1.根据实验室标准操作程序，将处理好的尿液样本放入生化分析仪器中。
2.设置所需参数（如波长、温度等），并启动仪器进行分析。
3.仪器会自动完成样本的检测和分析，并给出相应的结果。
6. 结果解读和记录1.根据尿微量蛋白分析的结果，判断尿液中蛋白质的含量。
2.根据临床标准，将结果归类为阴性、微量阳性、阳性等级。
3.将分析结果记录在相关表格或系统中，并进行必要的数据备份。
7. 质控和质量保证1.在实验过程中，使用已知浓度的标准品进行质控，以确保实验结果准确可靠。
尿液检验标准SOP
尿液标本采集和质量控制（基本常识和要求）1、尿液标本采集的一般要求1.1.患者告知：尿液标本采集，应首先告知患者关于尿液标本留取的目的，以口头和书面的形式具体指导尿液标本留取的方法。
1.2.病人一般准备的提示：1.2.1.患者应处于安静状态，按平常生活饮食。
1.2.2.用于细菌培养的尿标本须在使用抗生素治疗前采集，以有利于细菌生长。
1.2.3.运动、性生活、月经、过度空腹或饮食、饮酒、吸烟及姿势和体位等可影响某些检查的结果。
1.2.4.清洁外生殖器、尿道口及周围皮肤，女性患者应特别避免阴道分泌物或经血污染尿液。
1.2.5.如采用导尿标本或耻骨上穿刺尿标本，一般应由医护人员先告知患者及家属有关注意事项，然后由医护人员进行采集。
采集婴幼儿尿，应由儿科医护人员指导，用小儿专用尿袋收集。
1.3.唯一标识：在尿液采集容器和检验申请单上，应准确标记患者姓名、性别、年龄、留尿日期和时间、尿。
量、标本种类等信息，或以条形码做唯一标识。
1.4.容器及材料准备 :1.4.1.容器要求：应具备以下特点：1.4.1.1.一次性使用，材料与尿液成分不发生反应，洁净（菌落计数小于104CFU/L）、防渗漏。
1.4.1.2容积50～100 ml，圆形开口且直径至少4～5cm。
1.4.1.3.底座宽而能直立，有盖可防止倾翻时尿液溢出，如尿标本需转运，容器还应为安全且易于启闭的密封装置。
1.5.采集时段尿（如24h尿）:容器的开口更大，容积至少应达2～3L，且在盖不透光。
1.6细菌培养:尿标本容器应采用特制的无菌容器，对于必须储存2h以上才能检测的尿标本，最好北能使用无菌容器。
1.7儿科:患者尿液采集使用专用的清洁柔软的聚乙烯塑料袋。
尿液标本采集系统。
1.8．离心管:用于尿液沉渣检验的离心管应清洁、透明、带刻度，刻度上应至少标明10ml、1ml、0.2ml，容积应大于12ml，试管底部呈锥形或缩窄形。
试管口尽可能具有密封装置。
最好使用一次性玻璃离心管或不易破碎的塑料试管。
尿微量白蛋白(MAU)测定试剂盒(胶体金免疫层析法)产品技术要求贝尔
尿微量白蛋白（MAU）测定试剂盒（胶体金免疫层析法）适用范围：于体外定量测定人尿液中的尿微量白蛋白(MAU)含量。
1.1包装规格20人份/盒1.2 主要组成成分本试剂盒由MAU检测卡、干燥剂和滴管组成。
MAU检测卡由试纸条外壳与试纸条构成。
试纸条由样品垫、胶体金垫（喷有由胶体金标记的MAU单克隆抗体）、层析膜（T线包被有MAU单克隆抗体，C线包被有羊抗鼠IgG抗体）、吸水纸、衬垫构成。
检测卡为20人份/盒，干燥剂为1个/袋，滴管为20个/盒。
2.1 物理性状2.1.1 外观试剂盒各组分齐全、完整；包装袋应密封性好无破损；标签清晰；材料附着牢固，条宽应适应于卡壳且装配紧密。
2.1.2 膜条宽度膜条宽度应不低于4.0mm。
2.1.3 液体移行速度液体移行速度应不低于10mm/min。
2.2 空白检测限应小于5.0mg/L。
2.3 重复性用10mg/L尿微量白蛋白（MAU）参考品和100mg/L尿微量白蛋白（MAU）参考品各重复检测10次，其变异系数（CV）应不大于15%。
2.4 准确度将200.0mg/L尿微量白蛋白(MAU)参考品加入到尿微量白蛋白（MAU）含量5.0mg/L 正常人尿液参考品中，按照体积比1:9混合，对混合后样本进行检测，回收率应在85%～115%范围内。
2.5 线性线性范围为[5.0,200]mg/L，试剂盒的相关系数r应≥0.99。
2.6 批间差用3个批号试剂盒分别对10.0mg/L尿微量白蛋白（MAU）参考品和100.0mg/L 尿微量白蛋白（MAU）参考品各重复检测10次，则3个批号试剂盒之间的批间相对偏差（R）应不大于15%。
2.7 稳定性效期稳定性：2～30℃条件下放置有效期12个月后一个月内，检测物理性状、空白检测限、重复性、准确度、线性应符合2.1～2.6项的要求。
糖化血红蛋白尿微量白蛋白快速检测仪的标准操作程序
糖化血红蛋白尿微量白蛋白快速检测仪的标准操作程序一、目的：提供糖化血红蛋白和尿微量白蛋白（包括尿微量白蛋白、尿肌酐、白蛋白/肌酐比率三项参数）定量检测，其中糖化血红蛋白适用于糖尿病的检测和诊断，而尿微量白蛋白、尿肌酐、白蛋白/肌酐比率三项参数则适用于早期肾病的检测和诊断。
二、适用范围：糖尿病患者，糖尿病肾病患者，高血压肾病患者等。
三、定义：糖化血红蛋白：糖化血红蛋白是人体血液中红细胞内的血红蛋白与血糖结合的产物。
血糖和血红蛋白的结合生成糖化血红蛋白是不可逆反应，并与血糖浓度成正比，且保持120天左右，所以可以观测到120天之前的血糖浓度。
糖化血红蛋白的英文代号为HbA1c。
糖化血红蛋白测试通常可以反映患者近8～12周的血糖控制情况。
微量白蛋白尿：微量白蛋白尿是指在尿中出现微量白蛋白。
白蛋白是一种血液中的正常蛋白质，但在生理条件下尿液中仅出现极少量白蛋白。
微量白蛋白尿反映肾脏异常渗漏蛋白质。
四、职责规定齐宁宁负责本SOP的编写与修订规定董军梅负责本SOP的培训规定刘雪梅负责本SOP的执行五、程序：1.糖化血红蛋白的操作流程：①核对医嘱，评估患者一般情况，相关病情，既往病史。
②评估患者的心理状态，对健康知识的掌握情况及合作程度，告知其操作的目的和配合要点，取得合作。
③护士洗手、戴口罩，携用物至病人床旁。
④用75%酒精按照无菌原则消毒患者手指后采血，用采样针吸血至淀粉塞上方的横线处。
⑤将采血后的采样针卡入试剂盒内，将条形码面向右侧，在扫描槽内进行试剂盒扫描。
⑥按显示屏显示打开舱门，将试剂盒嵌入卡槽内，条码向右，缓慢而有力的拔出银色标签。
⑦在显示屏上填写患者信息。
⑧按开始按钮，测量开始，等待6分钟后自动打印出结果。
⑨打开舱门，拔出试剂盒，置于医用垃圾桶内，关机。
⑩洗手，记录结果。
2.尿微量白蛋白的操作流程①核对医嘱，评估患者一般情况，相关病情，既往病史。
②评估患者的心理状态，对健康知识的掌握情况及合作程度，告知其操作的目的和配合要点，取得合作。
尿微量白蛋白检测的标准操作程序
尿微量白蛋白检测的标准操作程序【目的】掌握NEPhstar特定蛋白分析仪检测尿微量白蛋白正确方法，保证检查项目的准确性。
【该SOP变动程序】本标准操作程序的改动，可由任一使用本SOP的工作人员提出，请专业组长及科主任签字后生效。
【检测原理】用散射比浊法，可溶性抗原与特异性的抗体反应形成不溶性复合物，当光线通过反应悬液时发生散射并由特种蛋白分析仪检测到。
散射光值的多少与测试样本中的蛋白浓度成一定比列。
通过刷卡将磁条中的标准曲线存储入仪器中，仪器会自动计算样品中的特定蛋白浓度。
【试剂组成】以上试剂由深圳国赛生物科技有限公司生产【所需其他材料】1．NEPHSTAR特定蛋白分析仪2．NEPHSTAR检测附件3．电子移液器YB2014．普通移液器【操作步骤】1、打开NEPHSTAR特定蛋白分析仪电源，仪器显示23、输入代码61，刷磁卡按刷磁卡，按ENTER直接进入下一步，默认仪器的序号和稀释度按此时4、将装有搅拌子和20ul尿液样品的测量杯放入测量室。
5移液器YB201将400ul缓冲液和40ul抗血清加入测量杯。
6、NEPHSTAR特定蛋白分析仪自动感应试剂的加入，搅拌子开始搅拌，测试开始。
测试完毕后仪器自动显示打印结果。
【参考区间】mALB<25mg/L【临床意义】1、正常情况下白蛋白在血液中检测出，并由肾脏过滤，当肾脏功能正常时白蛋白不会出现在尿液中，当肾小球基底膜受到损伤至通透性改变时，会有小部分白蛋白漏入尿液，形成微量白蛋白尿，高血压、糖尿病及系统性红斑狼疮等常伴有肾脏病变的缓慢进行性恶化，尿液中可较早发现异常。
2、尿液中白蛋白排泄量变动较大，随机尿标本一次尿微量白蛋白升高可能并无意义，如连续2-3次升高均超过参考区间方有诊断价值。
【注意事项】1、不要将批号的试剂要刷磁卡，磁卡里含有重要的标准曲线信息。
不同批号试剂不能混用。
2、试剂盒未开启前应存于2-8℃冰箱。
缓冲液和样品处理液在使用前放置室温平衡至18-26℃.抗血清和质控可稳定1个月。
cusabio 微量白蛋白尿(MAU ALB)检测试剂盒使用说明书
Human microalbunminuria(MAU/ALB) ELISA kit Catalog Number. CSB-E08970hFor the quantitative determination of human microalbunminuria(MAU/ALB) concentrations in serum, plasma, urine.This package insert must be read in its entirety before using this product.If You Have ProblemsTechnical Service Contact informationPhone: 86-27-87582341Fax: 86-27-87196150Email:****************Web: In order to obtain higher efficiency service, please ready to supply the lot numberof the kit to us (found on the outside of the box).1PRINCIPLE OF THE ASSAYThis assay employs the competitive inhibition enzyme immunoassay technique. Antibody specific for MAU has been pre-coated onto a microplate. Standards and samples are pipetted into the wells with biotin-conjugated MAU. A competitive inhibition reaction is launched between MAU (Standards or samples) and biotin-conjugated MAU with the pre-coated MAU antibody. After washing, avidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound reagent, a substrate solution is added to the wells and color develops in opposite to the amount of MAU bound in the initial step. The color development is stopped and the intensity of the color is measured.DETECTION RANGE0.078 µg/ml-5 µg/ml.SENSITIVITYThe minimum detectable dose of human MAU is typically less than 0.019 µg/ml. The sensitivity of this assay, or Lower Limit of Detection (LLD) was defined as the lowest human MAU concentration that could be differentiated from zero.SPECIFICITYThis assay has high sensitivity and excellent specificity for detection of human MAU. No significant cross-reactivity or interference between human MAU and analogues was observed.Note: Limited by current skills and knowledge, it is impossible for us to complete the cross-reactivity detection between human MAU and all the analogues, therefore, cross reaction may still exist.2PRECISIONIntra-assay Precision (Precision within an assay): CV%<8%Three samples of known concentration were tested twenty times on one plate to assess.Inter-assay Precision (Precision between assays):CV%<10%Three samples of known concentration were tested in twenty assays to assess.LIMITATIONS OF THE PROCEDUREFOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES.The kit should not be used beyond the expiration date on the kit label.Do not mix or substitute reagents with those from other lots or sources.If samples generate values higher than the highest standard, dilute the samples with Sample Diluent and repeat the assay.Any variation in Sample Diluent, operator, pipetting technique, washing technique, incubation time or temperature, and kit age can cause variation in binding.This assay is designed to eliminate interference by soluble receptors, binding proteins, and other factors present in biological samples. Until all factors have been tested in the Immunoassay, the possibility of interference cannot be excluded.3MATERIALS PROVIDEDReagents QuantityAssay plate (12 x 8 coated Microwells) 1(96 wells) Standard (Freeze dried) 2Biotin-conjugate (100 x concentrate) 1 x 60 µlHRP-avidin (100 x concentrate) 1 x 120 µlBiotin-conjugate Diluent 1 x 10 mlHRP-avidin Diluent 1 x 20 ml Sample Diluent 2 x 20 mlWash Buffer (25 x concentrate) 1 x 20 mlTMB Substrate 1 x 10 mlStop Solution 1 x 10 ml Adhesive Strip (For 96 wells) 4Instruction manual 1STORAGEUnopenedkitStore at 2 - 8°C. Do not use the kit beyond the expiration date.Opened kitCoated assayplateMay be stored for up to 1 month at 2 - 8°C.Try to keep it in a sealed aluminum foil bag,and avoid the damp.Standard May be stored for up to 1 month at 2 - 8° C.If don’t make recent use, better keep it storeat -20°C.HRP-avidinBiotin-conjugateBiotin-conjugateDiluentMay be stored for up to 1 month at 2 - 8°C. HRP-avidinDiluentSample DiluentWash BufferTMB SubstrateStop Solution*Provided this is within the expiration date of the kit.4OTHER SUPPLIES REQUIREDMicroplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.An incubator which can provide stable incubation conditions up to 37°C±0.5°C.Squirt bottle, manifold dispenser, or automated microplate washer.Absorbent paper for blotting the microtiter plate.100ml and 500ml graduated cylinders.Deionized or distilled water.Pipettes and pipette tips.Test tubes for dilution.PRECAUTIONSThe Stop Solution provided with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.5SAMPLE COLLECTION AND STORAGESerum Use a serum separator tube (SST) and allow samples to clot for30 minutes before centrifugation for 15 minutes at 1000 x g, 2 - 8°C.Remove serum and assay immediately or aliquot and store samples at -20°C or -80°C. Avoid repeated freeze-thaw cycles. Centrifuge the sample again after thawing before the assay.Plasma Collect plasma using EDTA, or heparin as an anticoagulant.Centrifuge for 15 minutes at 1000 x g, 2 - 8°C within 30 minutes of collection. Assay immediately or aliquot and store samples at -20°C or -80°C. Avoid repeated freeze-thaw cycles. Centrifuge the sample again after thawing before the assay.Urine Use a sterile container to collect urine samples. Remove any particulates by centrifugation for 15 minutes at 1000xg, 2 - 8°C and assay immediately or aliquot and store samples at -20°C or -80°C. Avoid repeated freeze-thaw cycles. Centrifuge again before assaying to remove any additional precipitates that may appear after storage.SAMPLE PREPARATIONRecommend to dilute the serum or plasma samples 50000-fold before test.The suggested 50000-fold dilution can be achieved by adding 2µl sample to 398µl of normal saline. Complete the 50000-fold dilution by adding 2µl of this solution to 498µl of Sample Diluent. The recommended dilution factor is for reference only. The optimal dilution factor should be determined by users according to their particular experiments.Recommend to dilute the urine samples with Sample Diluent(1:40) before test. The suggested 40-fold dilution can be achieved by adding 6µl sample to 234µl of Sample Diluent. The recommended dilution factor is for reference only. The optimal dilution factor should be determined by users according to their particular experiments6Note:1. CUSABIO is only responsible for the kit itself, but not for the samplesconsumed during the assay. The user should calculate the possible amount of the samples used in the whole test. Please reserve sufficient samples in advance.2. Samples to be used within 5 days may be stored at 2-8°C, otherwisesamples must be stored at -20°C (≤1month) or -80°C (≤2month) to avoid loss of bioactivity and contamination.3. Grossly hemolyzed samples are not suitable for use in this assay.4. If the samples are not indicated in the manual, a preliminary experiment todetermine the validity of the kit is necessary.5. Please predict the concentration before assaying. If values for these arenot within the range of the standard curve, users must determine the optimal sample dilutions for their particular experiments.6. Tissue or cell extraction samples prepared by chemical lysis buffer maycause unexpected ELISA results due to the impacts of certain chemicals.7. Owing to the possibility of mismatching between antigen from otherresource and antibody used in our kits (e.g., antibody targets conformational epitope rather than linear epitope), some native or recombinant proteins from other manufacturers may not be recognized by our products.8. Influenced by the factors including cell viability, cell number and alsosampling time, samples from cell culture supernatant may not be detected by the kit.9. Fresh samples without long time storage are recommended for the test.Otherwise, protein degradation and denaturalization may occur in those samples and finally lead to wrong results.7REAGENT PREPARATIONNote:Kindly use graduated containers to prepare the reagent. Please don't prepare the reagent directly in the Diluent vials provided in the kit. Bring all reagents to room temperature (18-25°C) before use for 30min.Prepare fresh standard for each assay. Use within 4 hours and discard after use.Making serial dilution in the wells directly is not permitted.Please carefully reconstitute Standards according to the instruction, and avoid foaming and mix gently until the crystals have completely dissolved.To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10µl for once pipetting.Distilled water is recommended to be used to make the preparation for reagents. Contaminated water or container for reagent preparation will influence the detection result.1. Biotin-conjugate (1x) - Centrifuge the vial before opening.Biotin-conjugate requires a 100-fold dilution. A suggested 100-fold dilution is 10 µl of Biotin-conjugate + 990 µl of Biotin-conjugate Diluent.2. HRP-avidin (1x) - Centrifuge the vial before opening.HRP-avidin requires a 100-fold dilution. A suggested 100-fold dilution is 10 µl of HRP-avidin + 990 µl of HRP-avidin Diluent.3. Wash Buffer(1x)- If crystals have formed in the concentrate, warm up toroom temperature and mix gently until the crystals have completely dissolved. Dilute 20 ml of Wash Buffer Concentrate (25 x) into deionized or distilled water to prepare 500 ml of Wash Buffer (1 x).894.StandardCentrifuge the standard vial at 6000-10000rpm for 30s.Reconstitute the Standard with 1.0 ml of Sample Diluent . Do not substitute other diluents. This reconstitution produces a stock solution of 5 µg/ml. Mix the standard to ensure complete reconstitution and allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making dilutions.Pipette 150 µl of Sample Diluent into each tube (S0-S6). Use the stock solution to produce a 2-fold dilution series (below). Mix each tube thoroughly before the next transfer. The undiluted Standard serves as the high standard (5 µg/ml). Sample Diluent serves as the zero standard (0 µg/ml).Tube S7 S6 S5S4 S3 S2 S1 S0 µg/ml52.51.250.6250.3120.1560.078ASSAY PROCEDUREBring all reagents and samples to room temperature before use. Centrifuge the sample again after thawing before the assay.It is recommended that all samples and standards be assayed in duplicate.1. Prepare all reagents, working standards, and samples as directed in theprevious sections.2. Refer to the Assay Layout Sheet to determine the number of wells to beused and put any remaining wells and the desiccant back into the pouch and seal the ziploc, store unused wells at 4°C.3. Set a Blank well without any solution.4. Add 50µl of standard and sample per well.5. Add 50µl of Biotin-conjugate(1x) to each well(not to Blank well). Coverwith a new adhesive strip. Incubate for 60 minutes at 37°C.(Biotin-conjugate(1x) may appear cloudy. Warm up to room temperature and mix gently until solution appears uniform.)6. Aspirate each well and wash, repeating the process two times for a total ofthree washes. Wash by filling each well with Wash Buffer (200µl) using a squirt bottle, multi-channel pipette, manifold dispenser, or autowasher, and let it stand for 2 minutes, complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.7. Add 100µl of HRP-avidin(1x) to each well(not to Blank well). Cover themicrotiter plate with a new adhesive strip. Incubate for 60 minutes at 37°C.8. Repeat the aspiration/wash process for five times as in step 6.9. Add 90µl of TMB Substrate to each well. Incubate for 20 minutes at 37°C.Protect from light.10. Add 50µl of Stop Solution to each well, gently tap the plate to ensurethorough mixing.1011. Determine the optical density of each well within 5 minutes, using amicroplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. Subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.*Samples may require dilution. Please refer to Sample Preparation section. Note:1. The final experimental results will be closely related to validity of theproducts, operation skills of the end users and the experimental environments.2. Samples or reagents addition: Please use the freshly prepared Standard.Please carefully add samples to wells and mix gently to avoid foaming. Do not touch the well wall as possible. For each step in the procedure, total dispensing time for addition of reagents or samples to the assay plate should not exceed 10 minutes. This will ensure equal elapsed time for each pipetting step, without interruption. Duplication of all standards and specimens, although not required, is recommended. To avoid cross-contamination, change pipette tips between additions of each standard level, between sample additions, and between reagent additions.Also, use separate reservoirs for each reagent.3. Incubation: To ensure accurate results, proper adhesion of plate sealersduring incubation steps is necessary. Do not allow wells to sit uncovered for extended periods between incubation steps. Once reagents have been added to the well strips, DO NOT let the strips DRY at any time during the assay. Incubation time and temperature must be observed.4. Washing: The wash procedure is critical. Complete removal of liquid ateach step is essential to good performance. After the last wash, remove any remaining Wash Solution by aspirating or decanting and remove any drop of water and fingerprint on the bottom of the plate. Insufficient washing will result in poor precision and falsely elevated absorbance reading. When using an automated plate washer, adding a 30 second soak period following the addition of wash buffer, and/or rotating the plate 180 degrees between wash steps may improve assay precision.115. Controlling of reaction time: Observe the change of color after adding TMBSubstrate (e.g. observation once every 10 minutes), TMB Substrate should change from colorless or light blue to gradations of blue. If the color is too deep, add Stop Solution in advance to avoid excessively strong reaction which will result in inaccurate absorbance reading.6. TMB Substrate is easily contaminated. TMB Substrate should remaincolorless or light blue until added to the plate. Please protect it from light.7. Stop Solution should be added to the plate in the same order as the TMBSubstrate. The color developed in the wells will turn from blue to yellow upon addition of the Stop Solution. Wells that are green in color indicate that the Stop Solution has not mixed thoroughly with the TMB Substrate.1213ASSAY PROCEDURE SUMMARY*Samples may require dilution. Please refer to Sample Preparation section.CALCULATION OF RESULTSUsing the professional soft "Curve Expert" to make a standard curve is recommended, which can be downloaded from our web.Average the duplicate readings for each standard and sample and subtract the average optical density of Blank.Create a standard curve by reducing the data using computer software capable of generating a four parameter logistic (4-PL) curve-fit. As an alternative, construct a standard curve by plotting the mean absorbance for each standard on the x-axis against the concentration on the y-axis and draw a best fit curve through the points on the graph. The data may be linearized by plotting the log of the MAU concentrations versus the log of the O.D. and the best fit line can be determined by regression analysis. This procedure will produce an adequate but less precise fit of the data.If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor.14人尿微量白蛋白(MAU/ALB)酶联免疫试剂盒使用说明书【产品编号】CSB-E08970h【预期应用】ELISA法定量测定人血清、血浆、尿液中MAU含量。
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其它适合的质控品
贮存条件：置2-8℃冰箱至有效期。
准备：直接使用。
质控间隔时间及限制：应视不同地区及各自实验室情况而定。质控结果应在限定的范围之内，如果超出范围，实验室应根据情况采取措施。
ABCD医院
生化实验室
文件编号：
ABCD-SOP-04-13
10.5 MAU的浓度低于3000mg/l时不会有HOOK效应。
ABCD医院
生化实验室
文件编号：
ABCD-SOP-04-13
尿微量白蛋白（MAU）测定
版序：ABCD
页码：第3页，共3页
11临床意义
白蛋白是非糖类蛋白，分子量为66，000道而顿。其在肝实质细胞中合成，每天大约为14g/天。白蛋白是血浆、脑脊液、尿液中最重要的蛋白成份（约大于50％）。尿液中少量的但是非正常的白蛋白排泌称为尿微量白蛋白。
<37mg albumin /g crea
7.2 24-hour尿液：<20mg/l
<30mg/24h
8性能指标
本法线性范围为3-120mg/dl，不准确度允许范围 ±10%，不精密度血液：CV=3.3%，尿液：CV=4.3%灵敏度为1.5mg/dl。
9注意事项
9.1血清标本出现溶血、脂血、黄疸或抗坏血酸的干扰情况参见抗干扰能力。
微量白蛋白尿可以由小球、小管及肾后性等原因引起。白蛋白是不同的类型的蛋白尿的标记蛋白。
对于选择性的肾小球性蛋白尿，排泌量约为100－3000mg/g crea。对于非选择性的肾小球性蛋白尿，其特点为高分子量蛋白的分泌增加。肾前性蛋白尿通过总蛋白与白蛋白之间的不同来确认（白蛋白少于30％）。白蛋白及微量蛋白同事增加可见于球管性蛋白尿（肾小管的重吸收少于肾小球的滤过率）。肾小球、肾小管以及间质性肾炎、糖尿病或其它原因引起的肾衰。血浆中的蛋白质具有两种功能：维持渗透压以及参与体内物质的转运。对于低水溶性物质来说，白蛋白是最重要的转运蛋白。白蛋白水平的降低可以由高血压、肝细胞合成减少、血管内分泌异常、血管内外分布异常，白蛋白丢失及降解加快等原因引起。
4.2校准物
来源：ROCHE配套校准物,符合WHO标准,CRM470，具体如下：
S1:0.9%NaCl
S2-6:定标液
贮存条件：如果防止细菌污染，可以2-8℃保存28天。
准备：直接使用。
定标频率：A试剂批号更换后
B由质控结果决定
4.3质控物
来源：Precinorm protein (罗氏蛋白正常值质控)
ABCD医院
生化实验室
文件编号：
ABCD-SOP-04-13
尿微量白蛋白（MAU）测定
版序：ABCD
页码：第1页，共3页
1测定方法
免疫比浊法。
2测定原理
尿液中微量的白蛋白（MAU）与试剂中包被有抗白蛋白抗体的胶乳颗粒相结合，与之发生凝集形成不溶性免疫复合物，使反应液产生混浊，MAU的浓度与胶乳颗粒的凝集形成的浊度成正比。
9.2换算公式：mg/dl×0.01=g/l
9.3仅应用于体外诊断。
9.4如果尿液的ALB的浓度偏高，应该应用NaCL,0.9%进行预稀释，建议的稀释比例如下：
ALB浓度
稀释比例
>300mg/l
1+1
>1000mg/l
1+10
>5000mg/l
1+ห้องสมุดไป่ตู้0
10抗干扰能力：
10.1标准：回收率在90％－110％之间。
3标本
血清及肝素-Li,Na,K/EDTA-K抗凝血浆，处理方法见标本准备。
稳定性：20–25℃7天
2 -8℃1个月
-20℃6个月
4试剂
4.1试剂
来源：ROCHE配套试剂（详见试剂说明书）。
贮存条件及稳定性：未打开试剂盒：2－8℃储存至效期末
R1：打开后机上稳定90天
R2：打开后机上稳定90天
准备：直接使用。
尿微量白蛋白（MAU）测定
版序：ABCD
页码：第2页，共3页
5仪器
ROCHE MODULAR P或日立7060生化分析仪。
6上机操作
见仪器作业指导书。
7参考范围
7.1 2nd晨尿：成年人：<20mg albumin/g crea
<2.26galbumin/mol crea
儿童（3-5岁）：<20mg/lalbumin
10.2黄疸：黄胆指数达到25时不会有明显干扰。（直接和间接胆红素浓度约为25mg/dl）
10.3溶血：溶血指数达到300时不会有明显干扰。（血红素浓度约为300mg/dl）
10.4 Vc<1g/l,Crea<5g/l,Glu<20g/l,UA<700mg/l,Urea<700mmol/l时不会造成明显干扰。