摘要：为实时监测种禽场及活禽市场的流感病毒流行情况，2016年8-12月对三家种禽场的种禽和南京周边地区活禽市场的家禽进行了禽流感病毒的分离和鉴定。共采集1110份咽拭子，进行核酸鉴定、病毒分离和亚型鉴定。结果表明：种禽场送检810份样本中，RT-PCR鉴定AIV阳性样本9份，其中H5亚型阳性3份，H7、H9亚型均为阴性，其余6份为其他亚型，阳性检出率为1.1%，阳性样品接种SPF鸡胚后未分离到病毒。活禽市场采集样本300份，RT-PCR检出AIV阳性样本共71份，其中H5阳性样本18份，H7阳性样本15份，其他亚型28份，阳性检出率23.70%。AIV阳性样本接种SPF鸡胚，72小时后血凝试验测定阳性19份，分离病毒株19份。本文为监测流感病毒流行病学和疫病防控预警奠定基础。28398
毕业论文关键词：禽流感病毒；分离鉴定；HA，亚型；序列分析
Isolation and identification of avain influenza A virus in farm and surrounding area of Nanjing
Abstract：In order to monitor the prevalence status of influenza virus in poultry farms and live poultry markets in real time, the avian influenza epidemiological survey of poultry, which from poultry markets and poultry farms, was conducted from August to December in 2016 and the virus was also isolated and identified.The pharyngeal swabs were inoculated into SPF chicken embryo. Collected Chicken embryonic allantoic fluid and subjected to hemagglutination after 72 hours. RT-PCR was used to isolate and identify positive samples. The genotypes of avian influenza virus were identified by H5, H7 and H9 subtypes, and some of the viruses were identified by HA gene sequencing. The results showed that there were 9 AIV positive samples detected in poultry farms, 3 of which was positive for H5, 0 for H7 and H9, 6 for the other subtypes. The positive rate was 1.1%.300 samples from the live poultry markets were isolated by RT-PCR . A total of 71 AIV positive samples were detected by RT-PCR. Among them, 18 were H5 positive samples, 15 were H7 positive samples and 28 were other subtypes. The positive samples were inoculated into SPF chichen embryo . We present 19 samples of hemagglutination test positive samples . The positive rate was 23.70%. This article lay the foundation for the detection of influenza virus epidemiology and disease prevention and control.
Keywords: avain influenza A virus; Isolation and Identification; HA; subtype; sequence analysis
目录
摘要 2
关键词： 2
Abstract： 2
Keywords: 2
1 材料与方法 3
1.1材料 3
1.1.1 咽拭子样本 3
1.1.2 SPF鸡胚 3
1.1.3试剂和菌株 3
1.1.4 仪器设备 3
1.1.5引物 3
1.2方法 4
1.2.1病毒的分离 4
1.2.2 病毒的血凝效价测定 4
1.2.3病毒RNA的提取 4
1.2.4病毒RNA的反转录 4
1.2.5 病毒的鉴定 4
1.2.6 病毒的分型 5
1.2.7病毒的测序 5
2结果与分析 6
2.1 样品RT-PCR检测 6
2.2病毒的分离鉴定 7
2.3 AIV分型鉴定 7
2.4 HA基因序列分析 7
3 讨论 8
致谢 10
参考文献： 10
附录A 10
种禽场及南京周边地区流感病毒的分离鉴定
禽流感病毒（avian influenza virus，AIV）属于正粘病毒科，是引发人类呼吸道疾病的主要原因[1]。禽流感病毒表面有血凝素（HA）蛋白和神经氨酸酶（NA）蛋白[2]。根据其抗原性的不同，可以将禽流感病毒分为16个HA亚型和9个NA亚型。根据其致病力的不同，可分为高致病性禽流感和低致病性禽流感。其中大部分高致病性禽流感为H5和H7亚型。 种禽场及南京周边地区流感病毒的分离鉴定:http://www.751com.cn/yixue/lunwen_23225.html