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

BMC Infectious Diseases

Kongxin Hu¹  Wei Zhen¹  Xiaohong Sun¹  Liping Zhang¹  Pengfei Yang³  Lei Shi²  Huanzhou Xu¹  Xuezheng Ma¹ 

[1] Institute of Health and Quarantine, Chinese Academy of Inspection and Quarantine,

[2] No.A3, Gaobeidian North Road, Beijing 100123, Chaoyang District, China;Department of Disease Control and Prevention, Shenzhen International Travel Health Care Center, Shenzhen 518045, Guangdong Province, China;Huaian Center for Disease Control and Prevention, No.118, Huaihai North Road, Qinghe District, Huaian, Jiangsu Province, China

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

Others  :  1135699
DOI  :  10.1186/s12879-015-0818-y

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

【 摘 要 】

Background

A 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.

Methods

The 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.

Results

Specificity 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.

Conclusions

The 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】

   
2015 Ma et al.; licensee BioMed Central.

【 图 表 】

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