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与传统蛋白质免疫印迹法相比,采用放射免疫蛋白质印迹法、第二代和第三代酶联免疫吸附测定法进行HIV-1感染的血清学诊断。

Radioimmune western blotting in comparison with conventional western blotting, second and third generation ELISA assays for the serodiagnosis of HIV-1 infection.

作者信息

Portincasa P, Conti G, Zannino T, Visalli S, Chezzi C

机构信息

Istituto di Microbiologia, Facoltà di Medicina e Chirurgia, Università degli Studi di Parma, Italy.

出版信息

New Microbiol. 1994 Jul;17(3):169-76.

PMID:7968651
Abstract

We compared the performance of second and third generation ELISA assays to detect antibodies to HIV-1 virus with conventional Western blotting (WB) and radioimmune Western blotting (RIWB). Both sera from commercial seroconversion panels and serial dilutions of a serum for HIV-1 antibodies were tested with Murex HIV Recombinant, Vidas bioMérieux HIV 1/2 (2nd generation ELISA) Murex HIV 1-2 (3rd generation ELISA), as well as with WB and RIWB. In seroconversion panels all ELISA assays were positive for the same serum with the exception of the first serum of Panel D which was negative with both sample Murex assays and borderline with Vidas assay. This serum was negative with WB but evidenced antibodies to gp160 p66, p51, p24 HIV-1 proteins when assayed by RIWB. In only two cases did WB reveal antibodies to HIV-1 proteins before ELISA assays (Panel A and E); not only did RIWB show the same sensitivity as WB in the two last panels, but it also detected antibodies to HIV-1 proteins earlier than WB, ranging from a few days (Panel C) to approximately 12 weeks (Panel D). The results obtained by testing the dilutions of the serum positive for anti HIV-1 antibodies showed the following degrees of sensitivity: Murex HIV 1-2 (the most sensitive), Murex HIV Recombinant and Vidas bioMérieux HIV 1/2. Although WB was more sensitive than the ELISA assays and picked out antibodies to gp160, gp120 and p24 HIV proteins at 1/4000 serum dilution, the most sensitive test was RIWB which at 1/20,000 serum dilution enabled detection of antibodies to gp160, p66 and p24 HIV proteins.

摘要

我们将第二代和第三代酶联免疫吸附测定(ELISA)检测HIV-1病毒抗体的性能与传统的蛋白质印迹法(WB)和放射免疫蛋白质印迹法(RIWB)进行了比较。使用Murex HIV重组检测法、生物梅里埃公司的Vidas HIV 1/2(第二代ELISA)、Murex HIV 1-2(第三代ELISA)以及WB和RIWB,对来自商业血清转化板的血清和一份HIV-1抗体血清的系列稀释液进行了检测。在血清转化板中,除了D组的第一份血清外,所有ELISA检测对同一血清均呈阳性,该血清在两种Murex检测样本中均为阴性,在Vidas检测中为临界值。该血清在WB检测中为阴性,但通过RIWB检测时显示出针对HIV-1病毒gp160、p66、p51、p24蛋白的抗体。仅在两例中,WB在ELISA检测之前就检测出了针对HIV-1蛋白的抗体(A组和E组);RIWB不仅在最后两组中显示出与WB相同的灵敏度,而且还比WB更早地检测出针对HIV-1蛋白的抗体,时间范围从几天(C组)到大约12周(D组)。通过检测抗HIV-1抗体阳性血清的稀释液所获得的结果显示出以下灵敏度程度:Murex HIV 1-2(最敏感)、Murex HIV重组检测法和生物梅里埃公司的Vidas HIV 1/2。尽管WB比ELISA检测更灵敏,并且在血清稀释至1/4000时就能检测出针对HIV病毒gp160、gp120和p24蛋白的抗体,但最灵敏的检测方法是RIWB,它在血清稀释至1/20,000时就能检测出针对HIV病毒gp160、p66和p24蛋白的抗体。

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