间接ELISA法检测钩端螺旋体特异性IgM方法的探讨

Establishment of an indirect ELISA for testing special IgM antibody of leptospira

为研制钩端螺旋体 (钩体 )特异性IgM检测试剂 ,并用于早期诊断钩体病 ,以钩体外膜蛋白为抗原 ,探索间接ELISA法检测钩体特异性IgM的各项实验条件 ,并对钩体病患者血清标本进行检测 ,结果 ,10 5份健康人血IgM阴性 ,钩体IgM阳性血清可被特异性阻断 ,2 巯基乙醇破坏试验阳性 ,试剂存放 4℃和 37℃ 4天的检测结果基本一致 ,检测临床钩体病人 112份血清 ,IgM阳性 83份 ,阳性率 74 11% ,与常规MAT法基本一致 ,0～ 7病日的阳性率明显高于MAT法。说明该法检测钩体病IgM具有特异、敏感、快速、稳定、简便等优点 ,对钩体病早期诊断有一定的价值。

In order to explore the method for testing special IgM against leptospira and apply it in the early diagnosis of the leptospria disease,all experimental parameters related to an indirect ELISA covered by the outer envelope protein of leptospira were studied.The blood samples of healthy students and the leptospira cases were tested by this method.The results showed that the IgM antibody against leptospira in 105 sera of healthy persons were negative,the specific IgM could be neutralized by the outer envelope protein of leptospira and destroied by 2 mercapto ethanol.The results were basically consistent in two detection kits stored at 4℃ and 37℃ for 4 days separatelly.Out of 112 sera of clinical leptospira cases,83 had positive IgM antibody,the positive rate was 74.11%,and it is similar to that of MAT method,but the former was higher significantly than the latter in 0～7 days of the disease.The results suggested this method is specific,sensitive,rapid,stable and simple,and had an obvious value during the early diagnosis of the leptospira patients.