Detection of hepatitis B virus DNA in serum samples via nested PCR and MALDI-TOF mass spectrometry

Genetic analysis, techniques and applications Pub Date : 1996-09-01 DOI:10.1016/1050-3862(95)00151-4

C. Jurinke , B. Zöllner , H.-H. Feucht , A. Jacob , J. Kirchhübel , A. Lüchow , D. van den Boom , R. Laufs , H. Köster

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引用次数: 26

Abstract

In a blind study, nested polymerase chain reaction (PCR) was performed with control DNA and DNA preparations from serum samples of six patients. The detection limit was determined to be 100 molecules of template in 1 ml of serum. Hepatitis B virus (HBV) related products of nested PCR were purified by ultrafiltration and immobilisation on streptavidin coated magnetic beads. The immobilized PCR products were denatured from the beads and analyzed via matrix assisted laser desorption/ionisation time-of-flight (MALDI-TOF) mass spectrometry. The results of MALDI-TOF MS analysis were in agreement with the results obtained by polyacrylamide gel electrophoresis (PAGE) and with the data obtained by serological analysis. The detection strategy introduced here has a high potential for automation and represents a fast and reliable method of detection for HBV DNA in serum without the need for time consuming gel electrophoresis and labeling or hybridization procedures.

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巢式PCR和MALDI-TOF质谱法检测血清乙型肝炎病毒DNA

采用巢式聚合酶链反应(PCR)对6例患者血清样本进行对照DNA和DNA制备。检测限为1 ml血清中100分子模板。采用超滤和链霉亲和素包被磁珠固定化的方法纯化巢式PCR的乙肝病毒相关产物。将固定PCR产物从微球上变性，并通过基质辅助激光解吸/电离飞行时间(MALDI-TOF)质谱分析。MALDI-TOF质谱分析结果与聚丙烯酰胺凝胶电泳(PAGE)和血清学分析结果一致。本文介绍的检测策略具有很高的自动化潜力，代表了一种快速可靠的血清HBV DNA检测方法，而不需要耗时的凝胶电泳和标记或杂交程序。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Genetic analysis, techniques and applications

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