Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Direct Multiplex Reverse Transcription-Nested PCR Detection of Influenza Viruses Without RNA Purification

  • Song, Man-Ki (Laboratory Science Division, International Vaccine Institute) ;
  • Chang, Jun (College of Pharmacy, Ewha Womans University) ;
  • Hong, Yeong-Jin (Department of Microbiology, Chonnam National University Medical School) ;
  • Hong, Sung-Hoi (Department of Clinical Laboratory Science, College of Health Sciences, Korea University) ;
  • Kim, Suhng-Wook (Department of Clinical Laboratory Science, College of Health Sciences, Korea University)
  • Published : 2009.11.30
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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.

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References

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