摘要
目的为了解人感染H7N9禽流感病毒基因特征从而更有效地防控疫情,需建立获得人感染H7N9禽流感病毒全基因序列的扩增和测序方法。方法采用鸡胚分离H7N9禽流感病毒,提取标本和毒株病毒RNA,自行设计引物,通过一步法RT-PCR扩增其全基因组片段,纯化PCR产物后测定核苷酸序列,利用生物信息学软件拼接各节段序列并初步分析其特征。结果本研究中的引物能特异地扩增各基因的目的片段,通过序列拼接可以获得H7N9禽流感全基因组序列。经分析8个节段序列与近期国内流行的人感染H7N9禽流感病毒序列高度同源。结论该方法能有效地获得人感染H7N9禽流感病毒全基因组序列,可为进一步分析其基因特征奠定基础。
There were three epidemic peaks of human infection with avian influenza A （H7N9） in China's Mainland in winter and spring since 2013. In this study, we established the assay of amplification and sequencing to obtain the complete genome of avian influenza virus A （HTN9）. After the RNA from chicken embryo-eggs isolated strains or throat swab specimen was ex- tracted, 22 fragments of genome were amplified by RT-PCR and sequenced with a set of self-designed primers. The complete genome sequence was obtained by sequence assembly and analyzed via bioinformatics. Each of the 8 genes of virus was closely related to that of HTN9 subtype viruses currently epidemic in China's Mainland. In conclusion, the assay could be used for ampli fying and sequencing the complete genome of avian influenza HTN9 subtype virus effectively, which could lay the foundation for the further analysis of gene characteristics.
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
2015年第9期795-799,共5页
Chinese Journal of Zoonoses
基金
国家科技重大专项(No.2013ZX10004104)
福建省医学创新课题(No.2014-CX-9)联合资助~~
关键词
H7N9禽流感
全基因组
扩增与测序
方法
avian influenza H7N9 subtype virus
complete genome
amplification and sequencing, methods
