Development of a multiplex real-time polymerase chain reaction for the detection of influenza virus type A including H5 and H9 subtypes.
Avian influenza viruses (AIVs) are endemic in wild birds and, if transmitted to poultry, can cause serious economic losses. The aim of this study was to develop a multiplex real-time reverse transcriptase polymerase chain reaction (RT-PCR) for rapid detection of influenza virus type A, including H5 and H9 subtypes. The selected primers and various labeled TaqMan reporter probes corresponding to matrix, H5, and H9 genes were used in a multiplex real-time RT-PCR to simultaneously detect triple fluorescent signals. The results showed that the multiplex real-time RT-PCR assay can be applied to detect RNA of influenza virus type A including H5 and H9 subtypes with a high specificity and a sensitivity of 10 copies per reaction. As a result of its short turnaround times and a high specificity and sensitivity, the assay is very suitable for large-scale screening during AIV outbreaks.
Li
Pei-Qiong
PQ
Department of Anatomy and DaAn Gene Diagnostic Center, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, Guangdong, PR China. lpq801105@yahoo.com.cn
Zhang
Jun
J
Muller
Claude P
CP
Chen
Jing-Xian
JX
Yang
Zi-Feng
ZF
Zhang
Ren
R
Li
Juan
J
He
Yun-Shao
YS
D000818:Animals; D001717:Birds; D017931:DNA Primers / Q000235:genetics; D005453:Fluorescence; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000235:genetics; D009980:Influenza A virus / Q000235:genetics / Q000302:isolation & purification*; D005585:Influenza in Birds / Q000175:diagnosis*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014763:Viral Matrix Proteins / Q000235:genetics
D017931:DNA Primers; D019267:Hemagglutinin Glycoproteins, Influenza Virus; C488936:M1 protein, Influenza A virus; D014763:Viral Matrix Proteins; C109963:hemagglutinin, avian influenza A virus