酶联免疫吸附试验检测人疱疹病毒7型抗体的研究

目的建立一种简便、快速、可靠的检测人疱疹病毒7型(HHV 7)特异性IgG和IgM酶联免疫吸附试验(ELISA)。方法应用HHV 7标准株G lasgow感染SupT1细胞,制备并纯化细胞工程抗原和重组pp85抗原;对325份血清HHV 7抗体进行检测,建立HHV 7特异性IgM和IgG间接ELISA,并与Q iagen公司ELISA试剂盒检测结果进行比较。结果采用细胞工程抗原和重组抗原制备的HHV 7 IgG和IgM ELISA诊断试剂敏感性、特异性、稳定性及重复性[变异系数(CV)<10%]较好;以Q iagen公司ELISA试剂盒为参比,重组抗原IgG ELISA检测上述标本的敏感性、特异性和符合率分别为98.1%、94.1%和97.8%,IgM ELISA分别为84.6%、99.7%和99.1%;与细胞工程抗原相比,重组抗原诊断试剂的特异性高而敏感性略低。结论自制HHV 7 IgG和IgMELISA检测试剂敏感特异,可用于临床HHV 7感染的诊断及流行病学调查。

Detection of antibody to human herpes virus 7 by enzyme-linked immunosorbent assay

Objective To establish a rapid,simple and reliable enzyme-linked immunosorbent assay(ELISA) for detection of IgG and IgM antibodies to human herpes virus 7(HHV 7). Methods SupT1 cells were infected with HHV 7(Glasgow) and harvested.Cellular antigen and recombinant antigen were produced and purified,indirect ELISA method for detection of IgG and IgM antibodies to HHV 7 was established,by coating the plate with the cell lysates and the recombinant proteins.It was used to screen 325 serum samples,which were simultaneously detected by an ELISA kit(from Qiagen Company). Results Our ELISA showed high specificity and sensitivity in the detection of HHV 7specific IgG and IgM antibodies in clinical samples.Compared with the ELISA kit produced by Qiagen Company,the sensitivity,specificity and coincident rate of our ELISA using recombinant protein for IgG were 98.1%,94.1% and 97.8% respectively,and that for IgM were 84.6%,99.7% and 99.1% respectively.The recombinant protein was more specific but less sensitive than the cells lysates. Conclusions The ELISA developed by our laboratory is sensitive and specific for the detection of IgG and IgM antibodies to HHV 7 and could be used for clinical diagnosis and epidemiology study.