Single-tube reverse transcription and heminested polymerase chain reaction of hepatitis C virus RNA to detect viremia in serologically negative hemodialysis patients
International Journal of Clinical & Laboratory ResearchPublished 1 March 1995
Maurizio Sampietro, Severo Salvadori, Noemi Corbetta, Salvatore Badalamenti, G. Graziani, G. Fiorelli
Citations11
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TL;DR
A substantial simplification of current methods for viral RNA reverse transcription and polymerase chain reaction amplification is described, which allows cDNA synthesis and nested amplification in a single tube through compartmentalization of the reagents for the two reactions with a wax barrier.
Abstract
Detection of hepatitis C virus genome is the only method currently available for detecting viremia in infected individuals. We describe a substantial simplification of current methods for viral RNA reverse transcription and polymerase chain reaction amplification, which allows cDNA synthesis and nested amplification in a single tube; this is achieved through compartmentalization of the reagents for the two reactions with a wax barrier. This procedure retains a high sensitivity: analysis of serum samples from hemodialysis patients, a high-risk group for hepatitis C virus infection, confirmed the presence of the virus in 88% of antibody-positive patients and detected viremia in patients without serological or biochemical evidence of hepatitis C virus infection.
Keywords
Agricultural and Biological SciencesMedicine
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