Coutlée F, Olivier C, Cassol S, Voyer H, Kessous-Elbaz A, Saint-Antoine P, He Y, Fauvel M
Centre de Recherche, Hôpital Notre-Dame, Montréal, PQ, Canada.
Am J Med. 1994 Jan;96(1):42-8. doi: 10.1016/0002-9343(94)90114-7.
The presence in some individuals of a prolonged phase of infection with human immunodeficiency virus type 1 (HIV-1) before seroconversion remains controversial. This study was undertaken to determine with a sensitive in vitro amplification technique, the polymerase chain reaction (PCR), whether seronegative individuals with high-risk behaviors could harbor HIV-1 sequences in their peripheral blood mononuclear cells (PBMCs) and remain seronegative for more than 6 months.
Seronegative individuals who engaged in unprotected anogenital intercourse with HIV-1-infected partners or with more than 10 individuals per year, and seronegative individuals who shared needles with seropositive partners, were recruited prospectively over 18 months. HIV-1 DNA and RNA sequences were detected in PBMCs of these individuals with three PCR assays using SK38/SK39, SK145/SK431, and SK68/SK69. Seronegative but PCR-positive patients were also evaluated with p24 antigen capture assay, radioimmunoprecipitation assay, and Western blot. The latter patients were followed prospectively to reproduce PCR-positive results and monitor serologic responses.
Sixty-one men and 18 women, with an average age of 34.1 +/- 7.6 years, were recruited: 56 were homosexual men, 18 were heterosexual women, and 5 were heterosexual men. Amplification reactions for HIV-1 of 104 PBMC specimens from 79 patients with negative or indeterminate serologies revealed that 4 patients (5.1%) were positive with PCR for HIV-1 DNA and RNA at the time of enrollment. Positive amplification reactions could not be reproduced in prospective samples for one patient. The analysis of a variable human genomic locus in this patient's PBMCs demonstrated that the first PCR-positive sample and following PCR-negative samples originated from different patients, suggesting a specimen mix-up. Two of the three PCR-positive seronegative patients had symptoms suggestive of acute retroviral disease. Sera from all three patients contained p24 antigen. Two patients seroconverted within 1 month whereas one patient could not be followed prospectively.
Prolonged infection with HIV-1 without seroconversion was not found in our population of patients at very high risk for HIV-1 infection. All PCR-positive patients seroconverted in less than 1 month.
在血清转化前,部分个体存在人类免疫缺陷病毒1型(HIV-1)感染的延长阶段,这一现象仍存在争议。本研究采用灵敏的体外扩增技术——聚合酶链反应(PCR),以确定具有高危行为的血清学阴性个体外周血单个核细胞(PBMC)中是否携带HIV-1序列,以及血清学阴性状态是否能持续超过6个月。
前瞻性招募血清学阴性个体,这些个体与HIV-1感染的性伴侣进行无保护的肛门生殖器性交或每年与超过10个人进行此类性交,以及与血清学阳性伴侣共用针头的血清学阴性个体,招募时间超过18个月。使用SK38/SK39、SK145/SK431和SK68/SK69三种PCR检测方法,在这些个体的PBMC中检测HIV-1 DNA和RNA序列。对血清学阴性但PCR阳性的患者,还采用p24抗原捕获试验、放射免疫沉淀试验和蛋白质印迹法进行评估。对后一类患者进行前瞻性随访,以重现PCR阳性结果并监测血清学反应。
共招募61名男性和18名女性,平均年龄34.1±7.6岁:56名是男同性恋者,18名是异性恋女性,5名是异性恋男性。对79例血清学阴性或不确定的患者的104份PBMC标本进行HIV-1扩增反应,结果显示4例患者(5.1%)在入组时HIV-1 DNA和RNA的PCR检测呈阳性。对一名患者的前瞻性样本未能重现阳性扩增反应。对该患者PBMC中一个可变的人类基因组位点进行分析表明,首个PCR阳性样本及随后PCR阴性样本来自不同患者,提示样本混淆。3例PCR阳性的血清学阴性患者中有2例有提示急性逆转录病毒病的症状。所有3例患者的血清中均含有p24抗原。2例患者在1个月内发生血清转化,而1例患者无法进行前瞻性随访。
在我们研究的HIV-1感染高风险人群中,未发现HIV-1的延长感染且无血清转化的情况。所有PCR阳性患者在1个月内均发生了血清转化。