摘要目的 初步建立针对非洲流行的病毒性出血热IgG抗体的多重检测方法.方法 重组表达并纯化非洲常见的出血热病毒抗原,将抗原偶联至荧光微球,单重检测兔血清IgG抗体评价偶联效果,多重检测从非洲入境的发热人员出血热病毒IgG抗体,并与ELISA方法比较,使用SPSS进行卡方检验和一致性分析.结果 重组表达纯化的出血热病毒抗原的纯度和浓度均达到偶联和检测要求.各重组抗原与相应荧光微球偶联有效,多重检测78份非洲入境发热人员血清样本,共检出1份拉沙热IgG抗体阳性、1份卢约病毒IgG抗体阳性,4份登革病毒IgG抗体阳性,6份黄热病毒IgG抗体阳性.与ELISA方法相比较结果无明显统计学差异,且两种方法具有高度相关性.结论 初步建立了基于Luminex高通量检测平台的针对非洲流行的病毒性出血热IgG抗体多重检测方法,为实现鉴别诊断提供了实验基础.

abstractsObjective To establish a multiplexed immunoassay for detection of IgG antibodies against viral hemorrhagic fever epidemic in Africa.Methods The recombinant antigens of hemorrhagic fever viruses (HFVs) epidemic in Africa were expressed and purified,and then coupled with the fluorescent microspheres.The coupling effects were evaluated by monoplexed detection of rabbit immune sera.Blood specimens were collected from people from Africa with fever,and multiplexed detection of IgG antibodies to HFVs was performed.Comparison of multiplexed assay and ELISA was performed by paired x2 test using SPSS software.Results Both the purity and concentration of HFVs recombinant antigen met the standards for coupling and detection,and the antigens were successfully coupled with fluorescent microspheres.Seventy-eight sera samples of people from Africa with fever were multiplex detected.Among these,one was tested positive for LASV-specific IgG,one was tested positive for LUJV-specific IgG,4 were tested positive for DENV-specific IgG and 6 tested positive for YFV-specific IgG.There was no statistically significant difference compared with ELISA,and the two method were highly correlated.Conclusions A multiplexed luminex-based immunoassay for detection of IgG antibodies to viral hemorrhagic fever epidemic in Africa was established,which laid the foundation for the differential diagnosis.