RAPID DETECTION OF GB-VIRUS-C RNA BY REVERSE TRANSCRIPTION-POLYMERASECHAIN-REACTION (RT-PCR) USING PRIMERS DERIVED FROM THE 5'NONTRANSLATED REGION

A simple reverse transcription-polymerase chain reaction (RT-PCR) proc edure for the detection of GB virus C (GBV-C) RNA in serum or plasma i s described. Ln this method, total nucleic acid, extracted from a smal l volume of human plasma, is reverse transcribed using random hexamers . An aliquot of cDNA is then utilized in PCR employing GBV-C specific primers designed to highly conserved regions of the 5'nontranslated re gion (NTR). For additional sensitivity, a second round of nested ampli fication is performed. Reactions are analyzed on an agarose gel and sa mples showing an ethidium bromide stained band of the appropriate size in the first and second amplification, or in the second amplification only, are designated to be positive. This protocol allows for the rap id and sensitive detection of GBV-C infection in human plasma or serum . (C) 1998 Elsevier Science B.V.