Quantification and duration of viraemia during hepatitis A infection as determined by real-time RT-PCR.
Human transmission studies and molecular techniques have provided evidence that transient viraemia occurs during infection with hepatitis A virus (HAV). However, the duration of its presence and levels during the phases of clinical disease and convalescence has not yet been well studied in human patients. Real-time RT-PCR techniques are increasingly used to quantify RNA viruses for diagnosis and/or research purposes. We have optimized a one-step RT-PCR that contains a dual-labelled fluorogenic probe to quantify the 5' noncoding region (5' NCR) of HAV. This method has a dynamic range (5-5 x 10(6) copies). The coefficient of regression of the standard curve was, on average 0.978. Intra-assay CVs% varied from 6.1% to 0.98%, and interassay CVs% from 6.46% to 2.1%. In the currently reported study 41 HAV IgM positive serum samples and 200 serum samples from healthy blood donors were tested by the quantitative RT-PCR method. The mean values on the first day of diagnosis found was 6.38 x 10(5) copies/mL. In a longitudinal study, viraemia persisted for an average of 60 days after clinical onset. These results show that viraemia in HAV infection lasts for many weeks.
Costa-Mattioli
M
M
Laboratoire de Virologie, Institut de Biologie, CHU Nantes, Nantes 44093, France.
Monpoeho
S
S
Nicand
E
E
Aleman
M-H
MH
Billaudel
S
S
Ferré
V
V
D000293:Adolescent; D000328:Adult; D002648:Child; D005260:Female; D006506:Hepatitis A / Q000821:virology*; D017957:Hepatitis A Virus, Human / Q000302:isolation & purification*; D006801:Humans; D008297:Male; D008875:Middle Aged; D012367:RNA, Viral / Q000097:blood; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D014766:Viremia / Q000821:virology*
D012367:RNA, Viral