APPLICATION OF NESTED PCR AND MASS-SPECTROMETRY FOR DNA-BASED VIRUS DETECTION - HBV-DNA DETECTED IN THE MAJORITY OF ISOLATED ANTI-HBC POSITIVE SERA

DNA preparations from three different groups of serum samples were exa mined for HBV-DNA via a nested polymerase chain reaction assay (lower detection limit: 10 viral genomes in 100 mu l serum): Group I consiste d of 11 uninfected control sera, group II consisted of sera obtained f rom 11 HBV infected patients and group III consisted of 21 isolated an ti-HBc positive samples. The 21 samples from group III were HBV-DNA ne gative according to a conventional non-nested PCR assay and hybridizat ion with a P-32-labelled probe. Using nested PCR and mass spectrometry , HBV-DNA was detected in none of group I and in all of group II sampl es. 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 H BV-DNA positivity and anti-HBc titers. Matrix-assisted laser desorptio n/ionization time-of-flight (MALDI-TOF) mass spectrometry provided a f ast, sensitive and non-radioactive assay for the detection of PCR prod ucts without the need for gel electrophoresis or hybridization with la belled probes. (C) 1998 Elsevier Science B.V.