肾综合征出血热IMMS-ELISA检测方法建立

目的建立一种用于检测肾综合征出血热(HFRS)特异性抗体IgG的ELISA方法—免疫磁性微球(IMMS)ELISA。方法利用重组菌株E.coli BL21(DE3)/pET32a-L99S诱导表达SEO型汉坦病毒(HV)重组核蛋白(rNP),并进行镍亲和层析纯化,以纯化rNP为抗原,分别建立检测HFRS特异性抗体IgG的3种ELISA法:间接ELISA、捕获ELISA和IMMS-ELISA,并进行方法学比较。结果新建立的3种ELISA的检测灵敏度和特异度为≥90%,总符合率≥95%。其中间接ELISA的灵敏度达100%,而假阳性率为10%;捕获ELISA特异度达100%,灵敏度较低(92.5%),且假阴性率为7.5%;IMMS-ELISA的灵敏度和特异度均达到100%。结论 IMMS-ELISA较其他2种ELISA检测方法更为简单、安全和准确,易于在基层公共卫生和临床医疗机构推广使用。

IMMS-ELISA :a test method for hemorrhagic fever with renal syndrome

Objective To establish a novel enzyme-linked immunosorbent assay(ELISA) method,namely immunomagnetic microspheres(IMMS)-ELISA,to detect specific IgG in hemorrhagic fever patients with renal syndrom(HFRS).Methods The recombinant nucleocapsid protein(rNP) of SEO hantavirus was expressed by the recombinant strain,E.coli BL21(DE3)/pET32a-L99S.Purified rNP was used as an antigen to establish three ELISA methods,including indirect ELISA,capture ELISA and IMMS-ELISA,for the detection of specific IgG in HFRS patients.The results of the three methods were compared with each other.Results The sensitivity and specificity of the three ELISA methods were all higher than 90% and their coincidence rates were all higher than 95%.The sensitivity of indirect ELISA was 100%,but its false positive rate was 10%.The specificity of capture ELISA was 100%,but its sensitivity was lower(92.5%) than that of other two methods.The sensitivity and specificity of IMMS-ELISA were all 100%.Conclusion IMMS-ELISA is a simpler,safer,and more accurate than indirect and capture ELISA method,and easy to use in grassroots public health and clinical medical institutions.