捕获法检测嗜肺军团菌IgM抗体的建立及应用  

DEVELOPMENT AND APPLICATION OF CAPTURE ELISA FOR DETECTING THE IGM OF LEGIONELLA PNEUMOPHILA

作　　者：万程彬[1] 

机构地区：[1]河南省南阳市第二人民医院,473012

出　　处：《中国煤炭工业医学杂志》2013年第9期1466-1468,共3页Chinese Journal of Coal Industry Medicine

摘　　要：目的建立一种捕获法检测嗜肺军团菌IgM抗体的方法。方法用羊抗人IgM抗体包被微孔板,选择辣根过氧化物酶标记的兔抗嗜肺军团菌抗体和培养的嗜肺军团菌抗原最适浓度,以西班牙VIRCELL公司PNEUMOSLIDE-M间接免疫荧光试剂盒为参照,优化样品稀释液和酶结合物稀释液的最佳配方。结果酶结合物工作浓度1/2 000、抗原浓度1/1 000为最适浓度,在样品稀释液中加入20%羊血清,在酶结合物稀释液中加入20%兔血清可以消除因使用多克隆抗体而引起的假阳性,假阳性率降低3.24%。建立的IgM捕获法与西班牙VIRCELL公司间接免疫荧光法的阳性符合率为93.26%(83/89),阴性符合率为93.67%(326/348),与其他呼吸道病原体IgM抗体没有显著交叉反应,不受类风湿因子的干扰。结论建立的捕获法检测嗜肺军团菌IgM抗体方法操作方便,与商品化间接免疫荧光试剂盒有较高的符合率,具有临床诊断价值。Objective To develop a capture ELISA for detecting the IgM of legionella pneumophila. Meth- ods The research used wells of microtiter plate coated with goat anti- human IgM McAb in the first step. Then the optimal concentration was chose for the reaction of horseradish peroxidase （HRP） labeled rabbit anti- legionella pneumophila McAb and the antigen of legionella pneumophila. Finally, the test was opti- mized for the sample dilution and enzyme conjugate dilution. The comparison method was used by indirect immunofluorescence Kit IPNEUMOSLIDE- M, VIRCELL Corporation of Spain}. Results The optimal working concentration of enzyme conjugate was 1/2 000, and that of antigen was 1/1 000. False positive that due to the using of polyclonal antibody can be eliminated through adding 20% goat serum in sample di- lution and 20% rabbit serum in enzyme conjugate dilution. Therefore, the false positive rate decreased 3.24%. The positive coincidence between capture ELISA method and the comparison method was 93.26% （83/89}, and the negative coincidence was 93.67% {326/348）. There was no significant cross reaction with others IgMs of respiratory passages pathogen. Otherwise, the capture ELISA wasn＇t interfered by rheumatoid factor. Conclusion The capture ELISA for detecting the IgM of legionella pneumophila is relia- ble, convenient and worth for clinical diagnose.

关 键 词：嗜肺军团菌 捕获法 IGM抗体 

分 类 号：R511.1[医药卫生—内科学]