This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Ravaggi, A. Articles by Cariani, E. Search for Related Content PubMed PubMed Citation Articles by Ravaggi, A. Articles by Cariani, E.

Next Article 

Journal of Clinical Microbiology, Feb 1995, 265-269, Vol 33, No. 2
Copyright © 1995 by the American Society for Microbiology. All rights reserved.

Quantification of hepatitis C virus RNA by competitive amplification of RNA from denatured serum and hybridization on microtiter plates

A Ravaggi, A Zonaro, C Mazza, A Albertini and E Cariani
Institute of Chemistry, School of Medicine, University of Brescia, Italy.

The direct detection of hepatitis C virus (HCV) RNA by PCR is widely used to determine the presence of circulating virions. The most relevant limit of this approach is the lack of quantitative information about the viral titer. We report a technique of competitive amplification allowing the estimation of HCV RNA copy number in biological samples. We constructed a standard competitive RNA template containing only two point mutations compared with its wild-type counterpart. The competitor was added in titrated amounts to the target RNA, and the mixture was then reverse transcribed and amplified in the same reaction tube. The relative amounts of target and competitor were determined by differential hybridization on microtiter plates with nonradioactive probes. The evaluation of HCV RNA titer required a single coamplification with the competitor and could be read from a standard curve. Furthermore, this method proved suitable for amplification of HCV RNA directly from serum, thus avoiding the intrinsic variability of the RNA extraction step.

This article has been cited by other articles:

• Preiser, W., Elzinger, B., Brink, N. S (2000). Quantitative molecular virology in patient management. J. Clin. Pathol. 53: 76-83 [Full Text]  
• Haberhausen, G., Pinsl, J., Kuhn, C.-C., Markert-Hahn, C. (1998). Comparative Study of Different Standardization Concepts in Quantitative Competitive Reverse Transcription-PCR Assays. J. Clin. Microbiol. 36: 628-633 [Abstract] [Full Text]