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地高辛标记的DNA探针:一种用于血清中乙肝病毒DNA检测的快速非放射性方法。

Digoxygenin-labelled DNA-probe: a rapid non-radioactive method for hepatitis B virus DNA detection in serum.

作者信息

Buti M, Jardi R, Rodriguez-Frias F, Arranz J A, Casacuberta J M, Sansegundo B, Puigdomenech P, Esteban R, Guardia J

机构信息

Department of Internal Medicine, Hospital Valle Hebrón, Universidad Autónoma, Barcelona, Spain.

出版信息

Eur J Clin Chem Clin Biochem. 1991 Nov;29(11):731-5. doi: 10.1515/cclm.1991.29.11.731.

Abstract

The sensitivity and specificity of two non-radioactive spot hybridization assays for hepatitis B virus DNA (HBV-DNA) using biotin and digoxygenin-labelled DNA probes were investigated in parallel in 122 serum samples from patients with chronic hepatitis B and 50 controls. The results were compared with an isotopic technique using a 32P-labelled probe. HBV-DNA was detected in 56 (80%) out of 70 hepatitis B "e" antigen (HBeAg)-positive cases and in 4 (8%) out of 52 antibody to hepatitis B "e" antigen (anti-HBe)-positive cases using the digoxygenin or 32P-labelled probes. No false positives were found with either method. Using the biotin-labelled probe, 16% of sera gave discordant results, which were considered to be false positive. The time required for detection of serum HBV-DNA was 2 hours for the non-radioactive probes and 16 hours for the isotopic probes. This study suggests that the digoxygenin-labelled probe for detection of HBV-DNA is the most rapid and sensitive method for routine diagnosis of viral replication in clinical laboratories.

摘要

使用生物素和地高辛标记的DNA探针的两种非放射性斑点杂交法检测乙型肝炎病毒DNA(HBV-DNA)的敏感性和特异性,在122份慢性乙型肝炎患者血清样本和50份对照样本中进行了平行研究。结果与使用32P标记探针的同位素技术进行了比较。使用地高辛或32P标记探针,在70例乙型肝炎e抗原(HBeAg)阳性病例中的56例(80%)以及52例乙型肝炎e抗体(抗-HBe)阳性病例中的4例(8%)检测到HBV-DNA。两种方法均未发现假阳性。使用生物素标记探针时,16%的血清给出不一致结果,被认为是假阳性。非放射性探针检测血清HBV-DNA所需时间为2小时,同位素探针为16小时。本研究表明,地高辛标记探针检测HBV-DNA是临床实验室常规诊断病毒复制最快速、灵敏的方法。

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