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首页> 外文期刊>Clinical and diagnostic laboratory immunology >Immune complex transfer enzyme immunoassay that is more sensitive and specific than western blotting for detection of antibody immunoglobulin G to human immunodeficiency virus type 1 in serum with recombinant pol and gag proteins as antigens.
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Immune complex transfer enzyme immunoassay that is more sensitive and specific than western blotting for detection of antibody immunoglobulin G to human immunodeficiency virus type 1 in serum with recombinant pol and gag proteins as antigens.

机译：以重组pol和gag蛋白为抗原的免疫复合物转移酶免疫测定法，比免疫印迹法更敏感，更特异性，可检测血清中针对1型人类免疫缺陷病毒的抗体免疫球蛋白G。

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Antibody immunoglobulin G (IgG) to human immunodeficiency virus type 1 (HIV-1) in serum was detected by ultrasensitive enzyme immunoassays (immune complex transfer enzyme immunoassays) with recombinant reverse transcriptase (rRT), p17 (rp17) and p24 (rp24) of HIV-1 as antigens and beta-D-galactosidase from Escherichia coli as the label. The immune complex, comprising 2,4-dinitrophenyl-bovine serum albumin-recombinant protein conjugate, antibody IgG to HIV-1, and recombinant protein-beta-D-galactosidase conjugate, was trapped on polystyrene beads coated with affinity-purified (anti-2,4-dinitrophenyl group) IgG, eluted with epsilon N-2,4-dinitrophenyl-L-lysine, and transferred to polystyrene beads coated with affinity-purified (anti-human IgG gamma-chain) IgG. Bound beta-D-galactosidase activity was assayed by fluorometry. The assays were highly reproducible with no serious serum interference, and they were much more sensitive than Western immunoblotting for the corresponding antigens. Signals with rRT, rp17, and rp24 for asymptomatic carriers were at least 56,000-, 680-, and 22-fold higher, respectively, than those for seronegative individuals, and neither indeterminate nor false-positive results were observed, whereas some serum samples were false negative or false positive by Western blotting for p17 and/or p24 antigen. In some cases, seroconversion was detected earlier than by conventional methods. Therefore, these assays are suggested to be more useful than conventional methods not only for the confirmation of antibody IgGs to RT, p17, and p24 of HIV-1 in serum but also for the detection of seroconversion.
机译：用超敏酶免疫测定法（免疫复合物转移酶免疫测定法），通过重组逆转录酶（rRT），p17（rp17）和p24（rp24）分别检测血清中针对人类1型免疫缺陷病毒（HIV-1）的抗体免疫球蛋白G（IgG）。以HIV-1作为抗原，以大肠杆菌的β-D-半乳糖苷酶为标记。免疫复合物被捕获在涂有亲和纯化的（抗-DNA抗体）的聚苯乙烯珠上，该复合物包含2,4-二硝基苯基-牛血清白蛋白重组蛋白结合物，抗HIV-1抗体IgG和重组蛋白-β-D-半乳糖苷酶结合物。 2,4-二硝基苯基）IgG，用εN-2,4-二硝基苯基-L-赖氨酸洗脱，并转移到涂有亲和纯化的（抗人IgGγ链）IgG的聚苯乙烯珠上。通过荧光法测定结合的β-D-半乳糖苷酶活性。该方法具有很高的重现性，没有严重的血清干扰，并且对相应抗原的敏感性比Western免疫印迹法高得多。无症状携带者的rRT，rp17和rp24信号分别比血清阴性个体至少高出56,000、680和22倍，并且未观察到不确定或假阳性结果，而一些血清样本通过Western印迹检测p17和/或p24抗原的假阴性或假阳性。在某些情况下，血清转化率比传统方法要早。因此，建议这些测定比常规方法更有用，不仅用于确认血清中HIV-1的RT，p17和p24的IgG抗体，而且还用于检测血清转化。

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