中国畜牧兽医2016，Vol.43Issue(12)：3101-3106,6.DOI:10.16431/j.cnki.1671-7236.2016.12.003
H9和 H6亚型禽流感病毒二重 RT-PCR检测方法的建立
Development of a Duplex RT-PCR Assay for Detection of H9 and H6 Subtype AIV
摘要
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 de-tection of H9 and H6 subtype AIV was developed by optimizing the PCR system such as the con-centration of different primers and annealing temperature.It showed that all samples could be am-plified specific bands from H9 subtype AIV single infection samples or H6 subtype AIV single in-fection 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 × 10 4 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.关键词
禽流感病毒/H9 亚型/H6 亚型/二重 RT-PCRKey words
avian influenza virus/H9 subtype/H6 subtype/duplex RT-PCR分类
农业科技引用本文复制引用
李丹,黄娇玲,谢芝勋,宋德贵,李孟,谢志勤,罗思思,谢丽基,黄莉,范晴..H9和 H6亚型禽流感病毒二重 RT-PCR检测方法的建立[J].中国畜牧兽医,2016,43(12):3101-3106,6.基金项目
国家“万人计划”领军人才专项(2016-37-88) （2016-37-88）
广西科技项目(2013GXNSFDA019015、AD163800009、14123001-8) （2013GXNSFDA019015、AD163800009、14123001-8）
广西水产畜牧局科研项目(桂渔牧科201204930、201452001) （桂渔牧科201204930、201452001）
