Loveday C, Tedder R S
Department of Medical Microbiology, University College and Middlesex School of Medicine, London, UK.
J Virol Methods. 1993 Feb;41(2):181-92. doi: 10.1016/0166-0934(93)90125-b.
Enzyme-linked immunosorbent assays (ELISA), using recombinant HIV-1 reverse transcriptase (RT; p66), are described for the measurement of RT antigen and serum antibodies to RT (anti-RT). The ELISA for anti-RT was developed in qualitative and quantitative forms, both were highly specific (100%, 0/859; 99.6%, 3/859), the former was sensitive (100%, 364/364) detecting the highest dilution of a standard high titre anti-HIV-1 RT antibody positive control serum. The latter was less sensitive (97.2%, 354/364) detecting lower dilutions of the antibody control, but had the advantage of producing highly reproducible optical density/concentration curves for the quantification of unknown anti-RT samples. In a cross-sectional study of 191 patients with HIV-1 infection, all patients developed anti-RT antibodies in CDC disease group II and III that declined but persisted in all cases into CDC disease group IV. The RT antigen assay was specific (100%, 0/772) and sensitive detecting 6 to 15 pg/ml of recombinant RT antigen diluted in normal human serum. No cross-reactivity using the RT antibody and antigen assays was seen in sera from 85 patients with current or previous hepatitis B infection or 21 sera from patients with HIV-2 infection. Further, no reactivity was demonstrated with the assays in a cohort of 20 seronegative partners (320 samples) exposed to HIV-1 infection over a 4-yr period. In samples from a patient with documented seroconversion, RT antigen was the first detectable marker of HIV-1 infection and was followed by a prompt anti-RT response. Serum RT antigen disappeared or remained low in most patients during CDC disease group II and III and rarely reappeared with progression to CDC disease group IV. In tissue culture studies RT antigen was detected in supernatant within 12 h (75 pg/ml), gave an initial peak at 36 h (300 pg/ml) and then continued to rise up to 5 days (603 pg/ml), offering a simple, cost-effective alternative to existing methods.
描述了使用重组HIV-1逆转录酶（RT；p66）的酶联免疫吸附测定（ELISA），用于测量RT抗原和抗RT血清抗体。抗RT的ELISA有定性和定量两种形式，两者特异性都很高（分别为100%，0/859；99.6%，3/859），前者敏感（100%，364/364），能检测到标准高滴度抗HIV-1 RT抗体阳性对照血清的最高稀释度。后者敏感性稍低（97.2%，354/364），能检测到抗体对照的较低稀释度，但优点是可为未知抗RT样品的定量产生高度可重复的光密度/浓度曲线。在一项对191例HIV-1感染患者的横断面研究中，所有患者在CDC疾病II组和III组中都产生了抗RT抗体，这些抗体下降但在所有病例中持续到CDC疾病IV组。RT抗原测定特异性高（100%，0/772），能敏感地检测到稀释在正常人血清中的6至15 pg/ml重组RT抗原。在85例现患或既往感染乙型肝炎患者的血清或21例HIV-2感染患者的血清中，使用RT抗体和抗原测定均未观察到交叉反应。此外，在一组20名在4年期间接触HIV-1感染的血清阴性伴侣（320份样本）中，该测定未显示反应性。在一名有血清转化记录的患者样本中，RT抗原是HIV-1感染的首个可检测标志物，随后迅速出现抗RT反应。在CDC疾病II组和III组期间，大多数患者的血清RT抗原消失或保持在低水平，在进展到CDC疾病IV组时很少再次出现。在组织培养研究中，RT抗原在12小时内可在上清液中检测到（75 pg/ml），在36小时出现初始峰值（300 pg/ml），然后持续上升至5天（603 pg/ml），为现有方法提供了一种简单、经济有效的替代方法。
