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KMID : 0545120090190111470
Journal of Microbiology and Biotechnology
2009 Volume.19 No. 11 p.1470 ~ p.1474
Direct Multiplex Reverse Transcription-Nested PCR Detection of Influenza Viruses Without RNA Purification
Song Man-Ki

Chang Jun
Hong Yeong-Jin
Hong Sung-Hoi
Kim Suhng-Wook
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.
KEYWORD
Influenza virus, multiplex RT-PCR
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