间接免疫荧光试验在HIV抗体检测中的应用

The Detection of HIV-1 Antibody by Indirect Immunofluorescence Assay (IFA)

[目的 ]评价间接免疫荧光试验 (IFA)作为一种确认实验在 HIV抗体检测中的应用。 [方法 ]用 T嗜性的HIV毒株感染 MT4细胞制备抗原片 ,并与不同稀释度的免疫血清相结合 ,再滴加荧光素标记的羊抗人 Ig G,在荧光显微镜下检测特异性免疫荧光。 [结果 ]用 IFA检测 43份血清样本 (其中包括 17份 WB阳性血清 ,14份阴性血清和 12份EL ISA阳性或 WB出现可疑条带但按标准不能判为阳性的可疑血清 ) ,除 HIV- 2血清呈阴性反应外 ,其余检测结果与WB相符。 IFA对 HIV- 1抗体检测的敏感性和特异性均达到 10 0 %。 HIV- 1阳性血清抗体最高滴度达到 1∶ 10 2 40。 [结论 ]IFA可用于 HIV- 1抗体检测的确认 ,它可作为 Western Blot的一种补充或替代实验。

Objective] To evaluate the application of IFA method as a confirmatory test for detecting the antibody of HIV. [Methods] HIV 1 infected MT4 cell suspension was spread on glass slides. Air dried cell smears were fixed in acetone. Tested sera were diluted and incubated with the antigen slide, and stained with fluorescein sothiocyanate conjugated goat anti human IgG, to detect the specific immunofluorescence. [Results] A panel of sera comprising 17 WB positive, 14 HIV antibody negative and 12 indeterminate sera (ELISA reactive but WB negative or indeterminate WB which followed up study showed negative for HIV 1 infection) were tested by IFA. Except for HIV 2 sera being negative, the IFA results of 42 specimens were equivalent relative to WB. The sensitivity and specificity of IFA was found to be 100%. The highest titers tested by IFA reached to 1:10240. [Conclusion] As a confirmatory test for HIV 1 antibody, IFA may be an alternative or supplementary test to Western Blot.