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Journal of Clinical Microbiology, June 2007, p. 1889-1892, Vol. 45, No. 6
0095-1137/07/$08.00+0     doi:10.1128/JCM.02392-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Human Influenza A Virus (H5N1) Detection by a Novel Multiplex PCR Typing Method{triangledown}

Shumei Zou,1 Jian Han,2 Leying Wen,1 Yan Liu,3 Kassi Cronin,2 Shanjuan H. Lum,2 Lu Gao,2 Jie Dong,1 Ye Zhang,1 Yuanji Guo,1 and Yuelong Shu1*

Chinese Center for Disease Control and Prevention, National Institute for Viral Disease Control and Prevention, State Key Laboratory of Infectious Disease Control and Prevention, Beijing, People's Republic of China,1 Genaco Biomedical Products, Inc., Huntsville, Alabama,2 Anhui Provincial Center for Disease Control and Prevention, Hefei, People's Republic of China3

Received 28 November 2006/ Returned for modification 19 January 2007/ Accepted 2 March 2007

We report the use of ResPlex III for genotyping influenza A viruses. The performance characteristics of the assay with regard to H5N1 are further evaluated. The ResPlex system incorporates a novel multiplex PCR technology, target-enriched multiplex PCR, to simultaneously amplify multiple molecular targets in one reaction. The ResPlex III assay targets the H1, H2, H3, H5, H7, H9, N1, and N2 genes from the influenza A virus as well as the NS genes from influenza A (NSA) and B (NSB) viruses, providing detection and genotyping of influenza A and B viruses. The analytical sensitivities for detecting the H5, N1, and NSA genes were 1, 10–1, and 10 50% tissue culture infectious doses/200 µl/reaction, respectively. A total of 217 sequential clinical samples including 14 samples with human H5N1 infections were tested by the ResPlex III assay, and the results were compared to a reference standard combined with results of viral culture and conventional reverse transcriptase and real-time PCR. The clinical sensitivity and specificity for detecting H5N1 were 93.3% and 100%, respectively, indicating that different subtypes of influenza A virus can be quickly and correctly identified using the ResPlex III genotyping approach.

* Corresponding author. Mailing address: China National Influenza Center, Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Yingxin Jie 100, Xuanwu District, Beijing 10052, People's Republic of China. Phone and Fax: 86-10-6357-7499. E-mail: yshu{at}vip.sina.com

{triangledown} Published ahead of print on 14 March 2007.

Journal of Clinical Microbiology, June 2007, p. 1889-1892, Vol. 45, No. 6
0095-1137/07/$08.00+0     doi:10.1128/JCM.02392-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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