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Reactivity patterns and infection status of serum samples with indeterminate Western immunoblot tests for antibody to human immunodeficiency virus type 1.

机译:用不确定的Western免疫印迹测试抗1型人类免疫缺陷病毒抗体的血清样品的反应性模式和感染状况

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摘要

Serum samples with indeterminate Western blot (WB) tests from 61 individuals whose sera were positive by enzyme-linked immunosorbent assay (ELISA) were studied in order to characterize their putative reactions with the human immunodeficiency virus (HIV) proteins and to resolve the HIV infection status of these individuals. The reaction observed by WB could not be confirmed either by radioimmunoprecipitation assay and subsequent electrophoresis (RIPA) or by use of LiaTek (Organon Teknika, Turnbout, The Netherlands) in 28% of the samples. Of the 86 samples that were indeterminate by WB, 66 reacted with p24 by WB; this reaction was confirmed by RIPA in only 21 (32%) and by LiaTek in 49 (74%) of the 66 samples. On the other hand, none of the indeterminate samples that reacted with HIV envelope proteins by WB did so by LiaTek, while 50% precipitated at least some of these proteins in the RIPA. The sensitivities of the three methods for detecting the antibody reaction with the different HIV proteins, which were studied with serial dilutions of positive serum samples, were similar. Thus, a lower sensitivity of RIPA or LiaTek does not seem to be the cause for the lack of reaction of the WB-indeterminate samples by these two methods. Sequential samples from individuals whose serum samples reacted by the three methods gave reproducible results, but all showed low antibody titers. Peripheral blood mononuclear cells obtained from three of the four individuals with sequential samples that reacted with HIV env proteins by WB and RIPA were negative for HIV provirus DNA after amplification by the polymerase chain reaction.
机译:研究了通过酶联免疫吸附测定(ELISA)对61份血清呈阳性的个体进行不确定的蛋白质印迹(WB)测试的血清样品,以表征其与人类免疫缺陷病毒(HIV)蛋白的假定反应并解决HIV感染这些人的地位。 WB观察到的反应无法通过放射免疫沉淀测定和随后的电泳(RIPA)或使用LiaTek(Organon Teknika,Turnbout,荷兰)在28%的样品中得到证实。在WB不确定的86个样品中,有66个样品与WB的p24反应; RIPA在66个样品中只有21个(32%)和LiaTek在49个(74%)中证实了该反应。另一方面,LiaTek并没有通过WB与HIV外壳蛋白反应的不确定样品,而50%的样品在RIPA中至少沉淀了其中一些蛋白。用连续稀释的阳性血清样品研究的三种检测与不同HIV蛋白反应的方法的灵敏度相似。因此,通过这两种方法,RIPA或LiaTek的较低灵敏度似乎并不是缺乏WB不确定样品反应的原因。来自血清样品通过三种方法反应的个体的顺序样品给出了可重现的结果,但所有抗体滴度均较低。从四名个体中的三名获得的外周血单核细胞经WB和RIPA与HIV env蛋白反应后,通过聚合酶链反应扩增后,其HIV原病毒DNA阴性。

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