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用于检测血清中乙肝病毒脱氧核糖核酸(HBV - DNA)的非放射性乙肝病毒脱氧核糖核酸探针

Non-radioactive hepatitis B virus DNA probe for detection of HBV-DNA in serum.

作者信息

Larzul D, Thiers V, Courouce A M, Bréchot C, Guesdon J L

机构信息

Laboratoire des Sondes Froides, Institut Pasteur, Paris, France.

出版信息

J Hepatol. 1987 Oct;5(2):199-204. doi: 10.1016/s0168-8278(87)80573-1.

Abstract

A diagnostic test has been developed to detect hepatitis B virus (HBV) DNA in human sera. This test involves a dot-blot technique in which non-radioactive nucleic acid labelled with 2-acetylaminofluorene (AAF) is used as probe. Two series of human sera from 228 blood donors and 113 HBsAg chronic carriers were tested by hybridization with the same DNA probe labelled either with AAF or with 32P. A correlation between the techniques was observed for 328 sera (96%), and using the non-radioactive test it was possible to detect 56 (86%) of the 65 HBV-DNA-positive patients. A comparative study of the HBeAg/anti-HBe status and the presence of HBV-DNA was carried out on the sera from 113 HBsAg chronic carriers, 65 of which were positive for HBeAg and 29 of which were positive for anti-HBe antibodies. With the AAF test, 44 of the HBeAg-positive sera were positive, while 5 of the anti-HBe-positive sera were positive. This study shows that, although this non-radioactive test is slightly less sensitive than the radioactive hybridization assay (RHA), it can be used for a survey of HBV carriers. Dissociation of the viral multiplication and the HBe/anti-HBe status was identified with the AAF test as well as with the RHA. It would therefore appear that the AAF test described here may be used for the extensive survey of HBV multiplication.

摘要

已开发出一种诊断测试来检测人血清中的乙型肝炎病毒(HBV)DNA。该测试采用斑点印迹技术,其中用2-乙酰氨基芴(AAF)标记的非放射性核酸用作探针。用标记有AAF或32P的相同DNA探针,通过杂交对来自228名献血者和113名HBsAg慢性携带者的两组人血清进行了检测。观察到328份血清(96%)的两种技术之间具有相关性,并且使用非放射性测试能够检测出65名HBV-DNA阳性患者中的56名(86%)。对113名HBsAg慢性携带者的血清进行了HBeAg/抗-HBe状态与HBV-DNA存在情况的比较研究,其中65名HBeAg呈阳性,29名抗-HBe抗体呈阳性。使用AAF测试,44份HBeAg阳性血清呈阳性,而5份抗-HBe阳性血清呈阳性。该研究表明,尽管这种非放射性测试的敏感性略低于放射性杂交分析(RHA),但它可用于HBV携带者的筛查。使用AAF测试以及RHA都发现了病毒增殖与HBe/抗-HBe状态的解离。因此,这里描述的AAF测试似乎可用于HBV增殖的广泛调查。

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