title: Direct multiplex reverse transcription-nested PCR detection of influenza viruses without RNA purification.
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.
authors.affiliation: Laboratory Science Division, International Vaccine Institute, Seoul 151-600, Korea.
authors.forename: Man Ki
authors.initials: MK
authors.lastname: Song
authors.forename: Jun
authors.initials: J
authors.lastname: Chang
authors.forename: Yeongjin
authors.initials: Y
authors.lastname: Hong
authors.forename: Sunghoi
authors.initials: S
authors.lastname: Hong
authors.forename: Suhng Wook
authors.initials: SW
authors.lastname: Kim
mesh_terms: D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D006358:Hot Temperature; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics / Q000302:isolation & purification; D053122:Influenza A Virus, H3N2 Subtype / Q000235:genetics / Q000302:isolation & purification; D009981:Influenza B virus / Q000235:genetics / Q000302:isolation & purification; D007251:Influenza, Human / Q000175:diagnosis* / Q000821:virology; D025202:Molecular Diagnostic Techniques / Q000379:methods; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012463:Saliva / Q000821:virology; D012680:Sensitivity and Specificity
chemical_list: D004279:DNA, Viral; D012367:RNA, Viral