Kestens L, Hoofd G, Gigase P L, Deleys R, van der Groen G
Institute of Tropical Medicine, Antwerpen, Belgium.
J Virol Methods. 1991 Jan;31(1):67-76. doi: 10.1016/0166-0934(91)90145-p.
Various methods were evaluated for their effectiveness in releasing HIV antigen (Ag) from artificial immune complexes (IC) and from IC present in serum from HIV antibody (Ab) positive subjects. The most effective methods for recovering HIV Ag from IC were those which included a denaturation step to prevent reassociation of Ag with Ab. IC precipitation in 2.5% polyethylene glycol followed by acid treatment with 1 M glycine.HCl (pH 2) for 10 min at 70 degrees C in the presence of 0.05% SDS gave very satisfactory results. With this method, IC were detected in sera from HIV antibody positive Caucasian subjects at all stages of infection. After HIV IC dissociation, HIV Ag was detected in a significant number (8/17 or 47%) of asymptomatic subjects. IC were most prevalent during the late stages of infection. A substantial increase in HIV Ag positivity was also observed in 20 Senegalese HIV Ab positive sera. After HIV IC dissociation HIV antigen detection increased from 2/20 to 12/20. The relevance of IC detection is discussed.
对多种从人工免疫复合物(IC)以及HIV抗体(Ab)阳性受试者血清中的IC释放HIV抗原(Ag)的方法进行了有效性评估。从IC中回收HIV Ag的最有效方法是那些包含变性步骤以防止Ag与Ab重新结合的方法。在0.05%十二烷基硫酸钠(SDS)存在的情况下,用2.5%聚乙二醇沉淀IC,随后在70℃下用1M甘氨酸盐酸盐(pH 2)进行酸处理10分钟,得到了非常令人满意的结果。用这种方法,在感染各阶段的HIV抗体阳性白种人受试者血清中均检测到了IC。HIV IC解离后,在大量(8/17或47%)无症状受试者中检测到了HIV Ag。IC在感染后期最为普遍。在20份塞内加尔HIV Ab阳性血清中也观察到HIV Ag阳性率大幅增加。HIV IC解离后,HIV抗原检测从2/20增加到12/20。文中讨论了IC检测的相关性。