A multiplex PCR assay for the detection of five influenza viruses using a dual priming oligonucleotide system

BMC Infectious Diseases

Research Article

Lei Shi¹  Pengfei Yang²  Huanzhou Xu³  Wei Zhen³  Xiaohong Sun³  Xuezheng Ma³  Liping Zhang³  Kongxin Hu³ 

[1] Department of Disease Control and Prevention, Shenzhen International Travel Health Care Center, 518045, Shenzhen, Guangdong Province, China;Huaian Center for Disease Control and Prevention, No.118, Huaihai North Road, Qinghe District, Huaian, Jiangsu Province, China;Institute of Health and Quarantine, Chinese Academy of Inspection and Quarantine,

[2] No.A3, Gaobeidian North Road, 100123, Beijing, Chaoyang District, China;

关键词: Dual priming oligonucleotide;    DPO;    Multiplex PCR;    Influenza;   

DOI  :  10.1186/s12879-015-0818-y

 received in 2014-08-04, accepted in 2015-02-10,  发布年份 2015

来源: Springer

【 摘 要 】

BackgroundA cost-effective, accurate and rapid simultaneous multiplex assay is required for testing and diagnoses of conventional and emerging viruses in clinical virology laboratories. We developed and optimized a dual priming oligonucleotide (DPO) multiplex PCR assay for detecting influenza viruses including seasonal H1N1, 2009 pandemic H1N1, H3N2, influenza B and H5N1.MethodsThe optimized multiplex DPO PCR was used to detect 233 clinical human samples. The results were compared to those obtained with RT-qPCR, conventional PCR and immunochromatographic assay.ResultsSpecificity analysis revealed that the DPO PCR assay amplified each target virus without any cross-amplification. Statistical analysis demonstrated that the multiplex DPO-PCR sensitivity was higher than for the immunochromatographic assay and lower than for qPCR, while no significant difference was observed compared with conventional PCR, when detecting influenza A and B. Additional experiments using the same sample panel indicated no significant differences between the number of positive samples detected by multiplex DPO PCR and RT-qPCR when applying a Cq with a value lower than 30.ConclusionsThe five-targeted simultaneous multiplex DPO PCR assay could be easily adopted into routine practice. This approach is cost effective with a short running time, low technical requirements for the detection of influenza virus and early diagnosis in clinical laboratories.

【 授权许可】

Unknown   
© Ma et al.; licensee BioMed Central. 2015. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.