Development of a Duplex RT-PCR Assay for Detection of H9 and H6 Subtype AIV |
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Authors: | LI DAN XIE Zhi-xun SONG De-gui LI Meng XIE Zhi-qin LUO Si-si XIE Li-ji HUANG Li FAN Qing HUANG Jiao-ling |
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Institution: | 1. College of Life Sciences, Guangxi Normal University, Guilin 541000, China;
2. Guangxi Key Laboratory of Vetrinary Biotechnology, Guangxi Veterinary Research Institute, Nanning 530001, China |
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Abstract: | This experiment was aimed to develop a method for simultaneous detection of H9 and H6 subtype avian influenza virus (AIV).Two pairs of specific primers were designed according to the conserved regions sequences of H6 and H9 AIV HA gene,a duplex RT-PCR simultaneous detection of H9 and H6 subtype AIV was developed by optimizing the PCR system such as the concentration of different primers and annealing temperature.It showed that all samples could be amplified specific bands from H9 subtype AIV single infection samples or H6 subtype AIV single infection samples,and the samples mix infection these two subtypes AIV.No specific bands of the same sizes were amplified from genomic materials of other avian pathogens.The detection limit of the duplex RT-PCR was 5×104 copies/μL. It suggested that this duplex RT-PCR assay was a specific,sensitive,stable and repeatable method for detection of H9 and H6 subtype of AIV,and could provide technical support for the monitoring of H9 and H6 subtype AIV. |
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Keywords: | avian influenza virus H9 subtype H6 subtype duplex RT-PCR |
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