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Single‐step nested polymerase chain reaction for detection of different genotypes of hepatitis C virus
Author(s) -
Li JiSu,
Tong ShuPing,
Vitvitski Ludmila,
Trépo Christian
Publication year - 1995
Publication title -
journal of medical virology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 121
eISSN - 1096-9071
pISSN - 0146-6615
DOI - 10.1002/jmv.1890450207
Subject(s) - nested polymerase chain reaction , virology , polymerase chain reaction , primer (cosmetics) , biology , hepatitis c virus , genotype , virus , reverse transcriptase , genetics , gene , chemistry , organic chemistry
Hepatitis C virus (HCV) exhibits considerable sequence variability and circulates in the blood at extremely low levels. Current methods for detecting HCV RNA are based mostly on nested polymerase chain reaction (PCR), in which part of the first amplification product is reamplified in the second tube by an internal primer pair. A novel nested PCR method was developed in which the two successive amplification processes are carried out in the same tube with a single step of physical manipulation. Careful selection of highly conserved sequences of the 5′ noncoding region as primers enabled successful detection of all three major genotypes circulating in France, including the one with variation in this region. Retaining high sensitivity of the conventional nested PCR, the novel method reduced greatly the risk of carry‐over contaminations. It was also cost‐ and time‐saving. The one‐step nested PCR method is especially suitable for routine diagnosis of HCV infection in clinical laboratories. © 1995 Wiley‐Liss, Inc.