Highly sensitive detection of viral RNA genomes in blood specimens by an optimized reverse transcription‐polymerase chain reaction

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Abstract
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.