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

作者：Ma, Xuezheng; Xu, Huanzhou; Shi, Lei; Yang, Pengfei; Zhang, Liping; Sun, Xiaohong; Zhen, Wei; Hu, Kongxin^*

来源：BMC Infectious Diseases, 2015, 15(1): 93.

DOI：10.1186/s12879-015-0818-y

• Dual priming oligonucleotide
• DPO
• Multiplex PCR
• Influenza

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.

• 出版日期2015-2-25
• 单位中国检验检疫科学研究院