Direct multiplex reverse transcription-nested PCR detection of influenza viruses without RNA purification

Man Ki Song, Jun Chang, Yeongjin Hong, Sunghoi Hong, Suhng Wook Kim

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

This paper describes the development a of direct multiplex reverse transcription-nested polymerase chain reaction (PCR) method, devised for simultaneous detection and typing of influenza viruses. This method combines the direct reverse transcription reaction without RNA purification with the enhancement of sensitivity and specificity of nested PCR. The method successfully detected three major human influenza viruses: influenza virus A subtype 1 (H1N1) and subtype 3 (H3N2), and influenza B virus (B). The minimum number of virus particles (pfu/ml) necessary for detection in spiked saliva samples was 200 (H1N1), 140 (H3N2), and 4.5 (B). The method's sensitivity and simplicity will be convenient for use in clinical laboratories for the detection and subtyping of influenza and possibly other RNA viruses.

Original languageEnglish
Pages (from-to)1470-1474
Number of pages5
JournalJournal of microbiology and biotechnology
Volume19
Issue number11
DOIs
Publication statusPublished - 2009 Nov

Keywords

  • Influenza virus
  • Multiplex RT-PCR

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology

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