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首页> 外文期刊>Journal of Clinical Microbiology >Specificities and sensitivities of three systems for determination of antibodies to human immunodeficiency virus by electrophoretic immunoblotting.
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Specificities and sensitivities of three systems for determination of antibodies to human immunodeficiency virus by electrophoretic immunoblotting.

机译：通过电泳免疫印迹测定人免疫缺陷病毒抗体的三种系统的特异性和敏感性。

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Electrophoretic immunoblotting (EIB [Western blotting]) has emerged as the major method for verification of seropositivity for human immunodeficiency virus (HIV) and therefore needs to be thoroughly characterized. The specificities of three EIB systems, our own and two commercial systems, were studied with anticellular sera and serial dilutions of human sera. We demonstrated that in one system, anti-HLA classes I and II gave bands comigrating with viral proteins, which can be controlled by EIB with uninfected H9 cells. In addition, animal antisera, including anti-immunoglobulin enzyme conjugates, occasionally reacted with HIV gag proteins, necessitating appropriate controls. Whereas none of 10 blood donors reacted at the standard dilution in serum (1/100 or 1/400) in any of the three systems, 6, 1, and 2 of 10 donors reacted with p24, p55, or both at a dilution of 1/10 for the three systems tested. Thus, nonspecific reactions can arise in several ways and justify critical EIB interpretation. The sensitivity of the three systems was studied by comparative titrations and direct quantification of bound immunoglobulin G (IgG). In the titrations with all three, the minor anti-HIV bands p53 and p64, coded from pol, were often detectable in higher dilutions than were antibodies to any other HIV protein. The minimum visible amounts of IgG bound per HIV protein band estimated by extra- and interpolation in densitometric curves and liquid scintillation counting of radiolabeled patient IgG were approximately 0.1, 0.05, and 0.02 ng per band in the three systems. One of the commercial systems had both the highest sensitivity and highest specificity.
机译：电泳免疫印迹（EIB [Western blotting]）已成为验证人免疫缺陷病毒（HIV）血清阳性的主要方法，因此需要进行全面鉴定。我们用抗细胞血清和人血清的系列稀释液研究了我们自己的三个商业EIB系统和两个商业系统的特异性。我们证明了在一个系统中，I类和II类抗HLA产生了与病毒蛋白竞争的条带，该条带可以由EIB用未感染的H9细胞控制。此外，动物抗血清，包括抗免疫球蛋白酶结合物，有时会与HIV gag蛋白反应，因此需要进行适当的控制。在三个系统中的任何一个系统中，有10个献血者中没有一个在血清中的标准稀释度（1/100或1/400）下发生反应，而10个献血者中的6个，1个和2个与p24，p55或两者都在稀释下反应了。测试的三个系统为1/10。因此，非特异性反应可以几种方式出现，并证明了关键的EIB解释是正确的。通过比较滴定法和结合免疫球蛋白G（IgG）的直接定量研究了这三种系统的敏感性。在用这三种方法进行滴定时，通常可以用比其他任何HIV蛋白抗体更高的稀释度检测到由pol编码的次要抗HIV带p53和p64。在三个系统中，通过光密度曲线的外加和内插以及放射性标记的患者IgG的液体闪烁计数估算的每个HIV蛋白条带结合的IgG的最小可见量约为每条带0.1 ng，0.05 ng和0.02 ng。一种商业系统具有最高的灵敏度和最高的特异性。

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