Richalet-Sécordel P M, Van Regenmortel M H
Laboratoire d'Immunochimie, Institut de Biologie Moléculaire et Cellulaire, Strasbourg, France.
FEMS Microbiol Immunol. 1991 Dec;4(1):57-64. doi: 10.1111/j.1574-6968.1991.tb04971.x.
A new capture test utilizing conjugated peptides has been developed for the detection of antibodies elicited against HIV-1. Human sera diluted 1:1000 were incubated in ELISA plates precoated with protein G. The captured IgG were allowed to react with three synthetic peptides corresponding to the gp41 sequence (591-611) YLKDQQLLGIWGCSGKLICTT, the gp120 sequence (314-329) IRIQRGPGRAFVTIGK and the p27 sequence (182-198) EWRFDSRLAFHHVAREL. The peptides were used in the form of N-hydroxysuccinimido-biotin ovalbumin conjugates. Peroxidase-labelled streptavidin was used to detect antigen-antibody complexes. The sensitivity and specificity of detection of antibodies were analyzed with 40 HIV positive sera, 10 seroconverting sera and 21 normal human sera (NHS). The results were compared with a commercial indirect ELISA in which a single conjugated gp41 peptide was used as antigenic probe. This indirect ELISA recognized 100% of the HIV positive and the seroconverting sera. The new capture test using the gp41 conjugated peptide also recognized 100% of the HIV positive sera but was more specific since it gave no false positive results whereas the indirect test did. The gp120 and p27 conjugated peptides detected 35/40 (87.5%) and 31/40 (77.5%) of HIV positive sera respectively and also detected 9/10 (90%) and 10/10 (100%) of the seroconverting sera respectively, without any false positive results (0/21). The proposed new capture test is a very sensitive and specific assay for detecting HIV antibodies.
已开发出一种利用偶联肽的新型捕获试验,用于检测针对HIV-1产生的抗体。将稀释至1:1000的人血清在预包被有蛋白G的ELISA板中孵育。使捕获的IgG与对应于gp41序列(591 - 611)YLKDQQLLGIWGCSGKLICTT、gp120序列(314 - 329)IRIQRGPGRAFVTIGK和p27序列(182 - 198)EWRFDSRLAFHHVAREL的三种合成肽反应。这些肽以N-羟基琥珀酰亚胺-生物素卵清蛋白偶联物的形式使用。过氧化物酶标记的链霉亲和素用于检测抗原-抗体复合物。用40份HIV阳性血清、10份血清转化期血清和21份正常人血清(NHS)分析抗体检测的敏感性和特异性。将结果与一种商业间接ELISA进行比较,该间接ELISA使用单一偶联的gp41肽作为抗原探针。这种间接ELISA识别出100%的HIV阳性和血清转化期血清。使用gp41偶联肽的新型捕获试验也识别出100%的HIV阳性血清,但更具特异性,因为它没有给出假阳性结果,而间接试验有。gp120和p27偶联肽分别检测出40份HIV阳性血清中的35份(87.5%)和31份(77.5%),也分别检测出10份血清转化期血清中的9份(90%)和10份(100%),且无任何假阳性结果(21份中为0份)。所提出的新型捕获试验是一种检测HIV抗体的非常敏感和特异的检测方法。