sharon sanders
Editor-in-Chief & President
doi:10.1016/j.diagmicrobio.2008.01.007 
Copyright ? 2008 Published by Elsevier Inc. Development of a multiplex real-time polymerase chain reaction for the detection of influenza virus type A including H5 and H9 subtypes<sup>
</sup>
Pei-qiong Li<sup>a</sup><sup>, </sup><sup>
</sup>, Jun Zhang<sup>a</sup>, Claude P. Muller<sup>b</sup>, Jing-xian Chen<sup>c</sup>, Zi-feng Yang<sup>c</sup>, Ren Zhang<sup>d</sup>, Juan Li<sup>a</sup> and Yun-shao He<sup>e</sup><sup>, </sup><sup>
</sup><sup>, </sup><sup></sup>
<sup>a</sup>Department of Anatomy and DaAn Gene Diagnostic Center, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, Guangdong, 510080 People's Republic of China
<sup>b</sup>Institute of Immunology, National Public Health Laboratory, L-1011 Luxembourg
<sup>c</sup>Virus Laboratory, Guangzhou Institute of Respiratory Disease, Guangzhou, Guangdong, 510120 People's Republic of China
<sup>d</sup>Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, 510407 People's Republic of China
<sup>e</sup>DaAn Gene Diagnostic Center, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, Guangdong, 510080 People's Republic of China
Received 4 November 2007; accepted 14 January 2008. Available online 4 March 2008.
Abstract
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.
Keywords: Avian influenza virus; Diagnose method; Multiplex real-time polymerase chain reaction
<sup></sup>All of the authors have no actual and potential conflict of interest for any persons and any organizations.
http://www.sciencedirect.com/scienc...serid=10&md5=1ed4fe5915e898a988f73b3855de6610
Copyright ? 2008 Published by Elsevier Inc. Development of a multiplex real-time polymerase chain reaction for the detection of influenza virus type A including H5 and H9 subtypes<sup>
</sup> Pei-qiong Li<sup>a</sup><sup>, </sup><sup>
</sup>, Jun Zhang<sup>a</sup>, Claude P. Muller<sup>b</sup>, Jing-xian Chen<sup>c</sup>, Zi-feng Yang<sup>c</sup>, Ren Zhang<sup>d</sup>, Juan Li<sup>a</sup> and Yun-shao He<sup>e</sup><sup>, </sup><sup></sup><sup>, </sup><sup></sup> <sup>a</sup>Department of Anatomy and DaAn Gene Diagnostic Center, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, Guangdong, 510080 People's Republic of China
<sup>b</sup>Institute of Immunology, National Public Health Laboratory, L-1011 Luxembourg
<sup>c</sup>Virus Laboratory, Guangzhou Institute of Respiratory Disease, Guangzhou, Guangdong, 510120 People's Republic of China
<sup>d</sup>Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, 510407 People's Republic of China
<sup>e</sup>DaAn Gene Diagnostic Center, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, Guangdong, 510080 People's Republic of China
Received 4 November 2007; accepted 14 January 2008. Available online 4 March 2008.
Abstract
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.
Keywords: Avian influenza virus; Diagnose method; Multiplex real-time polymerase chain reaction
<sup>
</sup>All of the authors have no actual and potential conflict of interest for any persons and any organizations.http://www.sciencedirect.com/scienc...serid=10&md5=1ed4fe5915e898a988f73b3855de6610