Study Of Influenza Viruses Nucleic Acid Detection Quality Control Materials Based On Recombinant MS2 Virus-like Particles

Influenza, leading to significant respiratory infections and resulting in 250,000 to 500,000 deaths worldwide annually, made it one of the most concerns to public health. With the expectation of timely epidemic evaluation, a national wide surveillance effort has been implemented to monitor influenza viruses.To evaluate the nucleic acid testes proficiency in influenza viruses (A/H1N1-2009, seasonal A/H1N1, A/H3N2, A/H5N1, A/H7N9, A/H9N2, B/Yamagata and B/Victoria) detection,37 types of different bacteriophage MS2 virus-like particles encapsulating full-length RNA sequences of Matrix protein, Nucleoprotein, non-structural protein, Hemagglutinin and Neuraminidase genes of different influenza viruses were constructed and expressed as influenza virus surrogate controls. Simultaneously,3 negative controls, for the purpose of probing the cross detection between different virus, were prepared by the bacteriophage MS2 virus-like particles that used in measles virus, rubella virus, and HCV external quality assessment programs. After confirming the copies of every bacteriophage MS2 virus-like particles and A/H3N2 by One-Step RT-ddPCR, the external quality assessment panel consisted of twenty-six influenza virus surrogate controls, one A/H3N2 virus and three negative controls was established.Though different kinds of commercial kits for nucleic acid extraction and TaqMan real-time RT-PCR kits for influenza virus RNA detection were employed by the 35 participants, the false positive rate of negative samples, false negative rate and misdiagnosis rate of positive samples are 0%(0/957),0.87%(14/1614) and 1.3% (21/1614). Overall, the panel used for influenza virus nucleic acid tests was successfully developed and the result of external quality assessment demonstrated that almost all of the participating laboratories possess a efficacious ability in influenza nucleic acid testes.