Automated quantitative determination of hepatitis C virus viremia by reverse transcription-PCR.
AUTOR(ES)
Besnard, N C
RESUMO
An automated reverse transcription-PCR was developed for the quantitative detection of hepatitis C virus. The quantitation is based on the coamplification and labelling with digoxigenin-dUTP during PCR of two similar templates, the viral genome and a modified RNA which acts as a mimic target. Known amounts of the mimic RNA sequence were introduced into the clinical samples. The automated quantitation of the two coamplified and labelled products depends on the use of two biotinylated caputre probes which are complementary, respectively, to a deleted sequence and to an inserted sequence introduced by site-directed mutagenesis in a wild viral cloned cDNA. This method proved to be simple, reproducible, and useful for quantitate hepatitis C virus viremia in chronically infected patients. This easy-to-perform, automated assay could also be used for the accurate determination of human immunodeficiency virus viremia or other RNA molecules.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=263897Documentos Relacionados
- Isolation of hepatitis C virus RNA from serum for reverse transcription-PCR.
- Detection of hepatitis E virus RNA in stools and serum by reverse transcription-PCR.
- Detection of African horse sickness virus by reverse transcription-PCR.
- Early diagnosis of Lassa fever by reverse transcription-PCR.
- Performance of the COBAS AMPLICOR HCV MONITOR Test, Version 2.0, an Automated Reverse Transcription-PCR Quantitative System for Hepatitis C Virus Load Determination