肾综合征出血热IMMS-ELISA检测方法建立  

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

作　　者：苏旭[1] 赵凤玲[2] 吕莉琨[1] 李力[1] 杨东靖[1] 陈锦英[3] 

机构地区：[1]天津市疾病预防控制中心,300011 [2]北京市通州区疾病预防控制中心 [3]天津医科大学

出　　处：《中国公共卫生》2012年第1期74-76,共3页Chinese Journal of Public Health

基　　金：天津市科技支撑计划重大项目(07SYSYSF05100)

摘　　要：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。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 meth- ods 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.

关 键 词：肾综合征出血热 汉坦病毒 免疫磁性微球 酶联免疫吸附测定 

分 类 号：R373.3[医药卫生—病原生物学]