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聚合酶链反应扩增后获得的人类免疫缺陷病毒前病毒DNA的临床相关性及基因多态性

Clinical correlation and genetic polymorphism of the human immunodeficiency virus proviral DNA obtained after polymerase chain reaction amplification.

作者信息

Genesca J, Wang R Y, Alter H J, Shih J W

机构信息

Department of Transfusion Medicine, Warren G. Magnuson Clinical Center, National Institutes of Health, Bethesda, MD 20892.

出版信息

J Infect Dis. 1990 Nov;162(5):1025-30. doi: 10.1093/infdis/162.5.1025.

DOI:10.1093/infdis/162.5.1025
PMID:2230230
Abstract

Polymerase chain reaction (PCR) was used to amplify a long sequence of human immunodeficiency virus (HIV) DNA, to assess the correlation between PCR signal and clinical stage of disease, and to demonstrate the genotypic variability of different HIV isolates. Twenty-four (96%) of 25 anti-HIV-reactive patients and none of 12 controls were positive for HIV proviral DNA by PCR. After quantification of the PCR signal, a significant difference in the relative amount of HIV proviral DNA per 10(5) peripheral blood mononuclear cells between symptomatic patients (Centers for Disease Control [CDC] class IV) (32,284 +/- 5225 cpm [mean +/- SE], equivalent to 802 HIV plasmid DNA copies) and patients without symptoms (CDC class II/III) (5484 +/- 1469 cpm [mean +/- SE], equivalent to 67 HIV plasmid DNA copies) was observed (P less than .01). Restriction analysis of PCR products in selected samples showed extensive genetic polymorphism between different isolates and more than one viral genotype per isolate. There was a clear correlation between the appearance of clinical symptoms in HIV infection and high levels of viral replication.

摘要

聚合酶链反应(PCR)用于扩增人类免疫缺陷病毒(HIV)DNA的长序列,以评估PCR信号与疾病临床阶段之间的相关性,并展示不同HIV分离株的基因变异性。25例抗HIV反应阳性患者中有24例(96%)通过PCR检测HIV前病毒DNA呈阳性,而12例对照均为阴性。对PCR信号进行定量后,有症状患者(疾病控制中心[CDC]IV级)每10⁵外周血单个核细胞中HIV前病毒DNA的相对量(32,284±5225 cpm[平均值±标准误],相当于802个HIV质粒DNA拷贝)与无症状患者(CDC II/III级)(5484±1469 cpm[平均值±标准误],相当于67个HIV质粒DNA拷贝)之间存在显著差异(P<0.01)。对选定样本中PCR产物的限制性分析显示,不同分离株之间存在广泛的基因多态性,且每个分离株有不止一种病毒基因型。HIV感染中临床症状的出现与高水平的病毒复制之间存在明显的相关性。

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