One-step multiplex reverse-transcriptase PCR for detecting pandemic (H1N1) 2009 influenza virus.
Pandemic (H1N1) 2009 influenza has spread throughout the world since April 2009 and has caused many human deaths since its first report in humans. Pandemic (H1N1) 2009 influenza virus was first identified in a Canadian pig herd in April 2009 and has been reported in more than ten countries, including Korea. We developed a one-step multiplex reverse transcriptase polymerase chain reaction (RT-PCR) assay based on the matrix gene that discriminates pandemic (H1N1) 2009 influenza virus from endemic swine influenza viruses. The sensitivity of this assay was 100 copies of in vitro-transcribed target RNA and 0.01 tissue culture infective dose (TCID(50)/ml) of virus and was as high as those of conventional influenza A virus common matrix reverse transcriptase PCR assays and real-time reverse transcriptase PCR assays (1 to 200 copies) developed for detecting pandemic (H1N1) 2009 influenza viruses from human and pig samples. This one-step multiplex RT-PCR assay would be a good tool in monitoring pandemic (H1N1) 2009 influenza virus among pig herds on a regular basis.
Shin
Yeun-Kyung
YK
Virology Division, National Veterinary Research and Quarantine Service, Ministry for Food, Agriculture, Forestry and Fisheries, Gyeonggi-do, Republic of Korea. shinyk@nvrqs.go.kr
Yoon
Soon-Seek
SS
Song
Jae-Young
JY
Kim
Joon-Bae
JB
Hang
Do Thi Thu
do TT
Park
Jin-Woo
JW
Kim
Min-Young
MY
Kim
Seong-Hee
SH
Park
Choi-Kyu
CK
Li
O-Soo
OS
D000818:Animals; D002460:Cell Line; D002642:Chick Embryo; D004285:Dogs; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000302:isolation & purification*; D007251:Influenza, Human / Q000453:epidemiology / Q000821:virology; D009976:Orthomyxoviridae Infections / Q000453:epidemiology / Q000662:veterinary* / Q000821:virology; D058873:Pandemics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013047:Specific Pathogen-Free Organisms; D013552:Swine