2016—2018年乌鲁木齐市某活禽市场禽流感病毒的分离与鉴定

目的 调查乌鲁木齐市活禽市场禽流感病毒（avain influenza virus，AIV）的流行情况。方法 2016年10月—2018年12月在该市某大型活禽市场采集鸡、鸭、鹅等活禽的双腔拭子1209份和环境样品75份，通过接种鸡胚、血凝抑制实验和AIVs PB1通用引物的RT - PCR等方法分离鉴定AIVs，以甲型流感病毒通用引物扩增病毒基因组8个片段并进行全基因组测序，利用BLAST和Clustal W等软件进行序列比对分析。结果 从90份样品中分离获得AIV，总的分离率为7.01%，其中家禽样品分离率为7.20%，鹅和鸭的分离率较高，分别为28.71%和13.01%，环境样品的分离率为4.00%。分离的AIV包括H5、H9、H3、H6、H1等多种亚型，其中H5和H9亚型病毒占比较高，均达到44.44%。AIV分离率呈明显的季节性变化，其中10—12月为高峰期。不同家禽及环境样品中AIV亚型及其分离率存在差异。结论 乌鲁木齐市活禽市场家禽和环境中存在多种亚型的AIV，急需加强对各类活禽市场的消毒与管理，降低AIV通过市场传播的风险。

Isolation and identification of avian influenza virus in a live poultry market in Urumqi from 2016 to 2018

To investigate the epidemic situation of avian influenza virus(AIV) in Urumqi live poultry market(LPM).Methods A total of 1 209 oropharyngeal and cloacal swabs from poultry(including chickens, ducks, and geese), as well as 75 environmental samples were collected from a large LPM in Urumqi during October 2016 to December 2018. AIVs were isolated and identified by inoculation of chicken embryo, hemagglutination inhibition test and RT-PCR with PB1 universal primer, the eight viral genome fragments were amplified with the universal primers of influenza A virus and the viral whole genome was sequenced, and pairwise sequence alignments analysis were performed by BLAST and Clustal W software.Results AIVs were isolated from 90 samples, the total isolation rate was 7.01%, the isolation rate of poultry samples was7.20%, the isolation rates of goose and duck were 28.71% and 13.01% respectively, and the isolation rate of environmental samples was 4.00%. The isolated AIVs included H5, H9, H3, H6, H1 and other subtypes, in which H5 and H9 subtypes accounted for 44.44%, respectively. The isolating rate of AIVs showed obvious seasonal variation, and October to December was the peak. The AIV subtypes and isolation rates were different in different poultry and environmental samples.Conclusion There are multiple AIV subtypes in the poultry and environment from Urumqi LPM. It is urgent to strengthen the disinfection and management of all kinds of LPMs to reduce the risk of AIV transmission through LPMs.