Single-tube reverse transcription and heminested polymerase chain reaction of hepatitis C virus RNA to detect viremia in serologically negative hemodialysis patients.
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.
Sampietro
M
M
Istituto di Medicina Interna e Fisiopatologia Medica, Università degli Studi di Milano, IRCCS Ospedale Maggiore Policlinico, Italy.
Salvadori
S
S
Corbetta
N
N
Badalamenti
S
S
Graziani
G
G
Fiorelli
G
G
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D005260:Female; D016174:Hepacivirus / Q000235:genetics* / Q000276:immunology; D006508:Hepatitis Antibodies / Q000097:blood; D006801:Humans; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000097:blood*; D012194:RNA-Directed DNA Polymerase; D006435:Renal Dialysis; D014766:Viremia / Q000175:diagnosis*
D006508:Hepatitis Antibodies; D012367:RNA, Viral; D012194:RNA-Directed DNA Polymerase