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

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• 作者：Xuezheng Ma (1)
  Huanzhou Xu (1)
  Lei Shi (2)
  Pengfei Yang (3)
  Liping Zhang (1)
  Xiaohong Sun (1)
  Wei Zhen (1)
  Kongxin Hu (1)
  
  1. Institute of Health and Quarantine ; Chinese Academy of Inspection and Quarantine ; |No.A3 ; Gaobeidian North Road ; Chaoyang District ; Beijing ; 100123 ; China
  2. Department of Disease Control and Prevention ; Shenzhen International Travel Health Care Center ; Shenzhen ; Guangdong Province ; 518045 ; China
  3. Huaian Center for Disease Control and Prevention ; No.118 ; Huaihai North Road ; Qinghe District ; Huaian ; Jiangsu Province ; China
  
• 关键词：Dual priming oligonucleotide ; DPO ; Multiplex PCR ; Influenza
• 刊名：BMC Infectious Diseases
• 出版年：2015
• 出版时间：December 2015
• 年：2015
• 卷：15
• 期：1
• 全文大小：1,893 KB
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• 刊物主题：Infectious Diseases; Parasitology; Medical Microbiology; Tropical Medicine; Internal Medicine;
• 出版者：BioMed Central
• ISSN：1471-2334

文摘

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.