IgM捕捉ELISA检测肾综合征出血热试剂盒研制

余鹏博; 王敬军; 张家驹; 郝志明; 刘中华; 胡兴菊; 唐青; 许文波

doi:10.11847/zgggws2006-22-02-29

IgM捕捉ELISA检测肾综合征出血热试剂盒研制

Study on IgM capturing ELISA kit for detecting hemorrhagic fever with renal syndrome

摘要

摘要:
目的开发可靠、稳定、方便的肾综合征出血热诊断试剂。
方法羊抗人μ链抗体包被聚苯乙烯酶标板, 纯化抗汉坦病毒核蛋白单克隆抗体, 并用辣根过氧化物酶标记后与纯化的汉坦病毒汉滩型(HTNV)核蛋白抗原一起使用作示踪剂, 结合四甲基联苯胺-过氧化氢(TMB/H₂O₂)酶底物系统, 制成检测肾综合征出血热早期特异性IgM抗体捕捉ELISA试剂盒。
结果与间接ELISA试剂同步检测46份出血热病人血清和46份正常人血清进行对比, 2种试剂检测结果完全相符, 特异性、稳定性和重现性好。
结论重组抗原IgM捕捉ELISA检测试剂盒敏感、可靠, 方便基层医院和门诊使用, 可用于肾综合征出血热的早期诊断及监测。

Abstract:
Objective To develop a sensitive, stable and simple diagnostic reagent for rapidly detecting epidemic hemorrhagic fever with renal syndrome(HFRS).
Methods The sheep anti-human μ chain was coated in polystyrene plate.The specific McAb of anti-nucleocapsid protein(NP)of Hantaan virus was purified and labeled with horseradish peroxidase (HRP).Recombined nucleocapside protein(rNP)antigen(Ag)was purified by chromato graphy.Both labeled McAb and rNP combined with enzyme substrate system(TMB/H₂O₂)play as a tracer.IgM capture ELISA kit of detecting anti-HFRS virus IgM was established.
Results Compaed with indirect ELISA, the total coincident rate of the results of simultaneous determination of 92 serum samples with epidemic hemorrhagic fever was 100%.The specificity, stability and repeatability of IgM capture ELISA kit were excellent.
Conclusion IgM capturing ELISA kit is sensitive, reliable and simple.It is applicable to hospitals and institutes detecting epidemic hemorrhagic fever with renal syndrome.It can be used for early diagnosis of epidemic hemorrhagic fever and epidemioligical studies.

HTML全文

参考文献(5)

施引文献

资源附件(0)