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血清中检测乙型肝炎病毒的DNA聚合酶反应产物分析——与斑点杂交技术的比较

Analysis of DNA polymerase reaction products for detecting hepatitis B virus in serum--comparison with spot hybridization technique.

作者信息

Imazeki F, Omata M, Yokosuka O, Matsuyama Y, Ito Y, Okuda K

出版信息

Hepatology. 1985 Sep-Oct;5(5):783-8. doi: 10.1002/hep.1840050513.

Abstract

An assay for DNA polymerase reaction products using slab gel electrophoresis and autoradiography was compared with the spot hybridization technique for the detection of hepatitis B virus DNA in 317 blood samples. The former could identify the nature and size of DNA on electrophoresis, and reduce potentially false-positive results due to artifacts. Discordant results between the two methods occurred in 36 of 317 samples; 22 were positive by the spot technique alone, and 14 were positive by the analysis of DNA polymerase reaction products alone. However, the samples positive with the spot test alone showed weak radioactive signals on electrophoresis/autoradiography that were often interpreted as "inconclusive" by blind observations. Correlation of hepatitis B e antigen/antibody with hepatitis B virus DNA was studied in 91 patients with various chronic liver diseases. Discordant results, i.e., presence of the DNA in antibody positive sera, or its absence in the antigen positive sera, were obtained in 15 (19%) cases. Such patients tended to have advanced liver disease with fluctuating serum aminotransferase levels. Analysis of DNA polymerase reaction products by slab gel electrophoresis and autoradiography is not only sensitive, but is also as specific as the Southern blot technique in the detection of hepatitis B virus DNA in serum, and may prove useful in selected samples, especially where no cloned hepatitis B virus DNA is available, or in search of new hepatitis B virus-like viruses.

摘要

采用平板凝胶电泳和放射自显影法对DNA聚合酶反应产物进行分析,并与斑点杂交技术进行比较,以检测317份血样中的乙型肝炎病毒DNA。前者可在电泳时鉴定DNA的性质和大小,并减少因假象导致的潜在假阳性结果。两种方法在317份样本中有36份结果不一致;22份仅斑点技术呈阳性,14份仅DNA聚合酶反应产物分析呈阳性。然而,仅斑点试验呈阳性的样本在电泳/放射自显影时显示出微弱的放射性信号,盲法观察时这些信号常被判定为“不确定”。对91例各种慢性肝病患者的乙型肝炎e抗原/抗体与乙型肝炎病毒DNA的相关性进行了研究。在15例(19%)患者中出现了不一致的结果,即抗体阳性血清中存在DNA,或抗原阳性血清中不存在DNA。这类患者往往患有晚期肝病,血清转氨酶水平波动。通过平板凝胶电泳和放射自显影法分析DNA聚合酶反应产物不仅灵敏,而且在检测血清中的乙型肝炎病毒DNA时与Southern印迹技术一样特异,在特定样本中可能有用,特别是在没有克隆的乙型肝炎病毒DNA的情况下,或用于寻找新的乙型肝炎病毒样病毒。

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