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Detection of the minus strand of hepatitis C virus RNA by reverse transcription and polymerase chain reaction: Implications for hepatitis C virus replication in infected tissue
Author(s) -
Takehara Tetsuo,
Hayashi Norio,
Mita Eiji,
Hagiwara Hideki,
Ueda Keiji,
Katayama Kazuhiro,
Kasahara Akinori,
Fusamoto Hideyuki,
Kamada Takenobu
Publication year - 1992
Publication title -
hepatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.488
H-Index - 361
eISSN - 1527-3350
pISSN - 0270-9139
DOI - 10.1002/hep.1840150305
Subject(s) - hepatitis b virus dna polymerase , hepatitis c virus , virology , virus , rna , hepatitis b virus pre beta , reverse transcriptase , biology , hepatitis b virus , ns2 3 protease , polymerase chain reaction , reverse transcription polymerase chain reaction , viral replication , hepatitis , microbiology and biotechnology , polymerase , dna , messenger rna , gene , biochemistry
Abstract The combination of reverse transcription and polymerase chain reaction is a very powerful tool for the detection of hepatitis C virus RNA in sera of patients with hepatitis C virus infection. However, when studying the presence of this virus in tissue using polymerase chain reaction, it may be difficult to distinguish between blood viral particles adhering to the tissue and viral RNA contained within the tissue. Because hepatitis C virus has a single‐stranded RNA of positive polarity, a minus‐strand RNA is expected to be found in hepatitis C virus–replicating tissues as a template for the synthesis of genomic RNA. To see whether the detection of the minus strand of hepatitis C virus RNA by polymerase chain reaction can be used for the determination of hepatitis C virus–replicating tissues, we examined the presence of the minus strand of hepatitis C virus RNA in the plasma, peripheral blood mononuclear cells and liver specimens of patients with hepatitis C virus infection. The plus‐strand RNA was detected in the plasma, peripheral blood mononuclear cells and the liver specimens, but the minus‐strand RNA was only detected in the liver. These results suggest that hepatitis C virus replicates in the liver but not in peripheral blood mononuclear cells. This detection method for the minus strand of hepatitis C virus RNA should be useful for determining hepatitis C virus replication in tissues other than liver tissue. (Hepatology 1992;15:387–390).