摘要
为了提高H9N2亚型禽流感病毒核酸检测结果的准确性,同时减少试验操作中的污染,对H9N2亚型禽流感病毒HA全长基因序列进行克隆及体外转录,构建用于检测H9N2亚型禽流感HA基因阳性核酸标准品。采用实时荧光定量RT-PCR方法对转录的RNA片段进行验证。结果表明:HA体外转录产物可以准确提供RNA片段的拷贝数并进行定性和定量分析。H9N2亚型禽流感病毒HA基因体外转录RNA可作为H9N2禽流感病毒核酸检测标准品的候选产物。
In order to improve the accuracy of nucleic acid detection results of H9 N2 subtype avian influenza virus and reduce the pollution in the experiment,the HA full-length gene sequence of H9 N2 subtype avian influenza virus was cloned and in vitro transcribed to construct a HA-positive nucleic acid standard for H9 N2 subtype avian influenza.The transcription RNA fragment was validated by real-time fluorescence quantitative RT-PCR.The results showed that the copy number of the RNA fragment could be accurately provided by the extracellular transcription product of HA,and qualitative and quantitative analysis was carried out.Therefore,in vitro transcription RNA of H9 N2 subtype avian influenza virus HA gene can be used as a candidate for the detection of H9 N2 avian influenza virus nucleic acid.
作者
李玥
孟庆峰
高明
姚贵哲
丛彦龙
王伟利
刘梦迪
LI Yue;MENG Qingfeng;GAO Ming;YAO Guizhe;CONG Yanlong;WANG Weili;LIU Mengdi(College of Animal Science and Technology,Jilin Agricultural University,Changchun 130118,China;Jilin Entry-Exit Inspection and Quarantine Bureau,Changchun 130000,China;College of Veterinary Medicine,Jilin University,Changchun 130062,China)
出处
《吉林农业大学学报》
CAS
CSCD
北大核心
2019年第2期231-235,共5页
Journal of Jilin Agricultural University
基金
吉林省科技攻关项目(20150204029NY)
关键词
H9N2亚型
禽流感病毒
体外转录
标准品
H9N2 subtype
avian influenza virus
in vitro transcription
standard
