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Microarray Multiplex Assay for the Simultaneous Detection and Discrimination of Influenza A and Influenza B Viruses
Authors:Mingyao Tian  Yufei Tian  Yang Li  Huijun Lu  Xiao Li  Chang Li  Fei Xue  Ningyi Jin
Institution:1. College of Animal Science and Veterinary Medicine, Jilin University, Changchun, 130062, People’s Republic of China
2. Institute of Military Veterinary, Academy of Military Medical Sciences of PLA, Liuying West Road 666, Changchun, 130122, People’s Republic of China
3. Department of Respiration, The First Hospital of Jilin University, Changchun, 130021, People’s Republic of China
Abstract:In this study, we present a microarray approach for the typing of influenza A and B viruses, and the subtyping of H1 and H3 subtypes. We designed four pairs of specific multiplex RT-PCR primers and eight specific oligonucleotide probes and prepared microarrays to identify the specific subtype of influenza virus. Through amplification and fluorescent marking of the multiplex RT-PCR products on the M gene of influenza A and B viruses and the HA gene of subtypes H1 and H3, the PCR products were hybridized with the microarray, and the results were analyzed using a microarray scanner. The results demonstrate that the chip developed by our research institute can detect influenza A and B viruses specifically and identify the subtypes H1 and H3 at a minimum concentration of 1 × 102 copies/μL of viral RNA. We tested 35 clinical samples and our results were identical to other fluorescent methods. The microarray approach developed in this study provides a reliable method for the monitoring and testing of seasonal influenza.
Keywords:Influenza A virus  Influenza B virus  Typing  DNA microarray
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