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Highly sensitive detection of viral RNA genomes in blood specimens by an optimized reverse transcription‐polymerase chain reaction
Author(s) -
Murakami Shigeki,
Takahashi Yoshiyuki,
Yoshida Shigeru,
Fuke Isao,
Ohmae Kozo,
Mori Chisato,
Takagi Mitsuo,
Takamizawa Akihisa,
Okayama Hiroto
Publication year - 1994
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.1890430215
Subject(s) - reverse transcriptase , virology , reverse transcription polymerase chain reaction , polymerase chain reaction , transcription (linguistics) , virus , rna , biology , nested polymerase chain reaction , microbiology and biotechnology , hepatitis c virus , polymerase , dna , gene , messenger rna , genetics , linguistics , philosophy
A protocol was developed for a highly sensitive detection of viral RNA in blood specimens by reverse transcription coupled with a nested poly‐merase chain reaction. Using Japanese encephalitis virus (JEV) as a model, the optimized reverse transcription‐polymerase chain reaction (ORTPCR) detects as few as 3–5 virions in 0.1 ml of whole blood specimens. The effectiveness of this assay system is confirmed by diagnosis of human hepatitis C viral (HCV) infection. © 1994 Wiley‐Liss, Inc.