Gaywee J, Artenstein A W, VanCott T C, Trichavaroj R, Sukchamnong A, Amlee P, de Souza M, McCutchan F E, Carr J K, Markowitz L E, Michael R, Nittayaphan S
Royal Thai Army, Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand.
J Acquir Immune Defic Syndr Hum Retrovirol. 1996 Dec 1;13(4):392-6. doi: 10.1097/00042560-199612010-00014.
The aim of this study was to compare the performance of differential polymerase chain reaction (PCR) typing and peptide enzyme-linked immunosorbent assay (V3-EIA) for human immunodeficiency virus type 1 (HIV-1) subtyping in Thailand using heteroduplex mobility assay (HMA) as the reference standard. Paired peripheral blood mononuclear cells (PBMC) and sera were collected from 38 HIV-1 seropositive persons in Thailand. HMA was done by standard methods; differential PCR employs primer pairs that differentially amplify either subtype E or B. V3-EIA used peptides specific for subtypes E or B. Thirty-two cases (84%) were found by HMA to be infected with subtype E: and six with (16%) subtype B. The results obtained with differential PCR were 100% concordant with those of HMA; V3 EIA correctly predicted the subtype in 95% (36 of 38). Six samples that molecularly subtyped as E were repeatedly dual reactive by screening V3-EIA, but these resolved to subtype E using an antigen-limiting EIA. Two samples were serologically nontypeable because of overall low levels of V3 antibody. Using HMA as the standard, differential PCR was shown to subtype HIV-1 reliably from patient PBMC samples. V3-EIA correctly predicted HIV-1 subtype in most (95%) of our cases. Because of the less rigorous sampling requirements, specimen processing, and logistical and technical requirements of serotyping compared with molecular techniques, it appears to be practical for screening purposes in a field environment. Samples that cannot be definitively subtyped serologically should undergo differential PCR and antigen-limiting V3 EIA. These approaches to HIV-1 subtyping should be used in complementary fashion in Thailand, where subtypes B and E are currently known to cocirculate.
本研究旨在以异源双链迁移率分析(HMA)作为参考标准,比较差异聚合酶链反应(PCR)分型和肽酶联免疫吸附测定(V3-EIA)在泰国对1型人类免疫缺陷病毒(HIV-1)进行亚型分型的性能。从泰国38名HIV-1血清阳性者中采集配对的外周血单核细胞(PBMC)和血清。HMA采用标准方法进行;差异PCR使用能分别特异性扩增E亚型或B亚型的引物对。V3-EIA使用针对E亚型或B亚型的特异性肽段。通过HMA发现32例(84%)感染E亚型,6例(16%)感染B亚型。差异PCR的结果与HMA的结果100%一致;V3-EIA在95%(38例中的36例)的样本中正确预测了亚型。6个分子分型为E亚型的样本在V3-EIA筛查中反复出现双反应性,但使用抗原限量EIA可确定为E亚型。2个样本由于V3抗体总体水平较低而无法进行血清学分型。以HMA作为标准,差异PCR被证明能从患者PBMC样本中可靠地对HIV-1进行亚型分型。V3-EIA在大多数(95%)病例中正确预测了HIV-1亚型。由于血清学分型与分子技术相比,采样要求、标本处理以及后勤和技术要求较宽松,因此在现场环境中用于筛查似乎是可行的。不能通过血清学明确亚型分型的样本应进行差异PCR和抗原限量V3-EIA检测。在目前已知B亚型和E亚型共同流行的泰国,这些HIV-1亚型分型方法应以互补的方式使用。