Design, development and evaluation of a competitive RT-PCR for quantitation of GBV-C RNA.
Although GB virus C (GBV-C) hepatocyte pathogenicity is still controversial, it appears that at least some strains of this virus are lymphotropic. During the past few years, several reports have documented an apparently beneficial role played by GBV-C in the course of HIV-1 infection. At present, a commercial kit for GBV-C RNA quantitation is not available. In this study, a competitive RT-PCR method for GBV-C in serum samples is described. The sensitivity of the assay proved to be 10(4) and 10(3) genomic equivalents for positive and negative sense RNAs, respectively. This method will discriminate specifically between positive and negative strand RNAs with a discrimination index of at least five log10. Out of 60 samples from different hematological disorders (n = 49), HIV-1 positive patients (n = 7), and blood donors (n = 4), 10 proved to be GBV-C RNA positive. Viral load ranged from 1.1 x 10(7) to 2.34 x 10(8) genomic equivalents/ml. Such values correlated linearly (r = 0.986) with those obtained by a 10-fold serial dilution method. In studies exploring the GBV-C pathogenicity, the measurement of viral load may contribute to understand the possible mechanisms involved.
Ruiz
Vanesa
V
Laboratorio de Hepatitis Virales, Departamento de Microbiología, Facultad de Medicina, Universidad de Buenos Aires, Paraguay 2155, Piso 11, 1121, Buenos Aires, Argentina. vanesruiz@yahoo.com
Espínola
Lidia
L
Mathet
Verónica Lidia
VL
Perandones
Carlos Edgardo
CE
Oubiña
José Raúl
JR
D029407:GB virus C / Q000302:isolation & purification*; D006801:Humans; D012367:RNA, Viral / Q000097:blood* / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D019562:Viral Load
D012367:RNA, Viral