用于检测和定量人类免疫缺陷病毒的灵敏逆转录酶检测法。
Sensitive reverse transcriptase assay to detect and quantitate human immunodeficiency virus.
作者信息
Lee M H, Sano K, Morales F E, Imagawa D T
机构信息
Department of Pediatrics, Harbor-University of California Los Angeles Medical Center, Torrance 90509.
出版信息
J Clin Microbiol. 1987 Sep;25(9):1717-21. doi: 10.1128/jcm.25.9.1717-1721.1987.
Abstract
A sensitive biochemical assay of viral reverse transcriptase (RT) was developed that is useful for both the detection and quantitation of human immunodeficiency virus (HIV), the agent responsible for acquired immune deficiency syndrome in humans. This assay gave a 20- to 40-fold increase in enzyme activity over the current method used for RT detection of HIV. The test is based on a previous biochemical study showing the unusual stability of avian oncornavirus RNA-dependent DNA polymerases at 30 degrees C for at least 2 days. Our study shows that the HIV polymerase is stable at 30 to 37 degrees C for up to 3 days. By using this sensitive RT assay, as few as 250 HIV virions can be quantitated directly in tissue culture medium. This assay should prove useful in studies in which the detection of HIV or the quantitation of the number of virions is required.
摘要
开发了一种灵敏的病毒逆转录酶（RT）生化检测方法，该方法对于人类免疫缺陷病毒（HIV）（即导致人类获得性免疫缺陷综合征的病原体）的检测和定量都很有用。与目前用于HIV逆转录酶检测的方法相比，该检测方法的酶活性提高了20至40倍。该测试基于先前的一项生化研究，该研究表明禽肿瘤病毒RNA依赖性DNA聚合酶在30℃下具有至少2天的异常稳定性。我们的研究表明，HIV聚合酶在30至37℃下可稳定长达3天。通过使用这种灵敏的逆转录酶检测方法，在组织培养基中可直接定量低至250个HIV病毒颗粒。该检测方法在需要检测HIV或定量病毒颗粒数量的研究中应会证明很有用。
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