Emmerich Petra, Thome-Bolduan Corinna, Drosten Christian, Gunther Stephan, Ban Enikö, Sawinsky Imke, Schmitz Herbert
Department of Virology, Bernhard-Nocht-Institute for Tropical Medicine, Bernhard-Nocht-Str. 74, D-20356 Hamburg, Germany.
J Clin Virol. 2006 Dec;37(4):277-81. doi: 10.1016/j.jcv.2006.08.015. Epub 2006 Sep 25.
Anti-Lassa antibodies are detected by indirect immunofluorescence assay (IFA) or by enzyme-immunoassay (ELISA). Both methods have problems to detect low amounts of specific antibodies.
We report here highly sensitive and specific reverse ELISAs to detect Lassa virus IgG and IgM antibodies. Due to the reverse techniques, serum samples could be applied at dilutions of 1:10 without increasing non-specific background reactions.
For IgM antibody detection microtiter plates were coated with anti-IgM antibodies and for IgG antibody detection with rheumatoid factor (RF) (Sachers M, Emmerich P, Mohr H, Schmitz H. Simple detection of antibodies to different viruses using rheumatoid factor and enzyme-labelled antigen (ELA). J Virol Methods 1985;10:99-110). In both assays a tissue culture antigen was used in combination with a labeled anti-Lassa monoclonal antibody (Hufert FT, Ludke W, Schmitz H. Epitope mapping of the Lassa virus nucleoprotein using monoclonal anti-nucleocapsid antibodies. Arch Virol 1989;106(3-4):201-12).
The reverse ELISA turned out to detect virus-specific IgG and IgM antibody in all 20 samples of West African patients collected 2-8 weeks after onset of Lassa fever. Moreover, both IFA and reverse ELISA found IgG antibodies in 53 out of 643 samples of healthy West Africans (sensitivity of 100%). Six of the 643 samples were positive by reverse IgG ELISA only. Thus, the specificity compared to IIF was 99.0%, but it may be even higher, because compared to IFA the IgG ELISA was clearly more sensitive in detecting low antibody titers.
In Ghana 3% seropositives were found by IFA, but 4% by the reverse ELISA. The reverse ELISAs can be performed with high sensitivity and specificity under field conditions in Africa.
抗拉沙病毒抗体通过间接免疫荧光法（IFA）或酶免疫法（ELISA）检测。两种方法在检测低水平特异性抗体时都存在问题。
我们在此报告用于检测拉沙病毒IgG和IgM抗体的高灵敏度和特异性的反向ELISA。由于采用了反向技术，血清样本可以以1:10的稀释度应用，而不会增加非特异性背景反应。
用于IgM抗体检测的微量滴定板用抗IgM抗体包被，用于IgG抗体检测的用类风湿因子（RF）包被（萨赫斯M、埃默里希P、莫尔H、施密茨H。使用类风湿因子和酶标记抗原（ELA）简单检测针对不同病毒的抗体。《病毒学方法杂志》1985年；10:99 - 110）。在这两种检测中，组织培养抗原与标记的抗拉沙单克隆抗体联合使用（胡费特FT、路德克W、施密茨H。使用单克隆抗核衣壳抗体对拉沙病毒核蛋白进行表位作图。《病毒学档案》1989年；106(3 - 4):201 - 12）。
反向ELISA在拉沙热发病后2 - 8周采集的所有20份西非患者样本中均检测到病毒特异性IgG和IgM抗体。此外，IFA和反向ELISA在643份健康西非人的样本中均检测到53份IgG抗体（灵敏度为100%）。643份样本中有6份仅通过反向IgG ELISA呈阳性。因此，与间接免疫荧光法相比特异性为99.0%，但可能更高，因为与IFA相比，IgG ELISA在检测低抗体滴度时明显更灵敏。
在加纳，IFA检测到3%的血清阳性，但反向ELISA检测到4%。反向ELISA在非洲现场条件下可以高灵敏度和特异性地进行。
