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Application of nested PCR and mass spectrometry for DNA-based virus detection: HBV-DNA detected in the majority of isolated anti-HBc positive sera.

作者信息

Jurinke C, Zöllner B, Feucht H H, van den Boom D, Jacob A, Polywka S, Laufs R, Köster H

机构信息

Department of Biochemistry and Molecular Biology, Faculty of Chemistry, University of Hamburg, Germany.

出版信息

Genet Anal. 1998 Jan;14(3):97-102. doi: 10.1016/s1050-3862(97)10006-7.

DOI:10.1016/s1050-3862(97)10006-7
PMID:9526701
Abstract

DNA preparations from three different groups of serum samples were examined for HBV-DNA via a nested polymerase chain reaction assay (lower detection limit: 10 viral genomes in 100 microliters serum): Group I consisted of 11 uninfected control sera, group II consisted of sera obtained from 11 HBV infected patients and group III consisted of 21 isolated anti-HBc positive samples. The 21 samples from group III were HBV-DNA negative according to a conventional non-nested PCR assay and hybridization with a 32P-labelled probe. Using nested PCR and mass spectrometry, HBV-DNA was detected in none of group I and in all of group II samples. In 11 out of 21 (52%) of the isolated anti-HBc positive sera from group III, HBV-DNA was detected. No correlation was observed between HBV-DNA positivity and anti-HBc titers. Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry provided a fast, sensitive and non-radioactive assay for the detection of PCR products without the need for gel electrophoresis or hybridization with labelled probes.

摘要

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