Abstract

PEREA, Yenitse; ARMAS, Anny; GONZALEZ, Yaimé J  and  LAZA, Celia M. Stability of standard curves of hepatitis C virus transcripts used for viral quantification. Biotecnol Apl [online]. 2009, vol.26, n.3, pp.222-225. ISSN 1027-2852.

The World Health Organization currently estimates the prevalence of hepatitis C virus (HCV) infections to be around 3%, representing approximately 170 million of infected people worldwide. An important step in the confirmatory diagnosis of HCV is the detection of its ribonucleic acid (RNA) in human serum or plasma, using nucleic acid amplification methods. Likewise, the measurement of HCV RNA viral loads by these methodologies is important for monitoring the therapeutic efficacy of antiviral drugs and monitor disease progression in HCV-infected individuals. The aim of this work was to study the stability of external curves built with different concentrations of an HCV transcript, which are used as components of a competitive Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) assay for the quantification of HCV RNA in human serum or plasma samples. The curves were studied for a period of 18 months by quantification at each time point against a fresh external curve, using an internal standard diluted in lysis buffer. RNA was extracted from each point of the curve and amplified by competitive RT-PCR, using a fluorimetric detection scheme based on the use of separate, specific probes for the HCV amplicons or the internal quantification standard. The results obtained in this work demonstrated that the RNA from each point of the quantification curve remained stable for all studied time points, allowing the use of ready-to-use curves, prepared in lysis buffer containing the internal quantification standard, for HCV viral load assays.

Keywords : RNA; HCV; quantification.

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