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http://dx.doi.org/10.4014/jmb.0905.05012

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)
Publication Information
Journal of Microbiology and Biotechnology / v.19, no.11, 2009 , pp. 1470-1474 More about this Journal
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.
Keywords
Influenza virus; multiplex RT-PCR;
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Times Cited By Web Of Science : 1  (Related Records In Web of Science)
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