Objective: To analyze the characteristics of spread and genetic evolution of H5 subtype avian influenza virus in Guangzhou from 2014 to 2019. Methods: H5 subtype virus was detected by fluorescence quantitative RT-PCR from the environmental samples in Guangzhou poultry markets. The genes of HA and NA of 48 isolates randomly selected were sequenced, including 46 isolates from environmental samples and 2 isolates from cases. The characteristics of molecular variation and genetic evolution were analyzed by using bioinformatics software. Results: A total of 1 094 strains of H5 subtype avian influenza virus were isolated from 52 284 samples (2.09%). All the strains belonged to Clade 2.3.4.4.C. NA gene belonged to H6N6 of Eurasian lineage. The cleavage sites of all the strains showed the characteristics of highly pathogenicity. Receptor binding sites were avian-derived receptors. However, mutations of S123P, S133A and T156A occurred, which implied that these strains could tend to bind to human receptors. There was an additional glycosylation site at 140 in strains isolated after 2017. The variation of antigen loci mainly occurred in B and E regions. Conclusions: H5 subtype avian influenza virus spread in Guangzhou from 2014 to 2019 with annual increased proportion of positive rate, and the sequencing results indicated that it belonged to Clade 2.3.4.4.C of H5N6 highly pathogenic virus, and genetic evolution and mutation continued, especially the common mutations which could enhance the binding capacity to human receptors. It is necessary to strengthen the surveillance.
目的： 分析广州市H5亚型禽流感病毒流行特点及基因进化和变异特征，为禽流感的防控提供依据。 方法： 对2014－2019年广州市禽类市场外环境标本进行禽流感病毒核酸检测并分析H5亚型流行情况，随机选取48份（46份来源于环境和2份来源于病例）H5阳性标本进行HA和NA基因测序，应用生物信息学软件分析分子遗传特征。 结果： 2014－2019年监测的52 284份环境标本中，检出H5亚型阳性标本1 094份，阳性率为2.09%。遗传进化分析显示，HA基因属于Clade 2.3.4.4.C分支，NA基因主要归属于欧亚谱系H6N6进化分支，HA和NA基因的进化以时间聚类为特点，相近年份流行株聚成一个进化分支。分子特征显示，48份毒株裂解位点呈现高致病性分子特点，受体结合位点仍为禽源受体，但普遍发生S123P、S133A和T156A倾向结合人源受体的位点突变。抗原位点变异主要出现在B、E区，并表现时间分布的特点，同一年份流行株常发生与上一年份流行株不同的抗原位点变异。2017年开始所有毒株出现由氨基酸缺失导致的糖基化位点的增加（140-NHT）。 结论： 广州市禽类市场外环境H5亚型禽流感病毒长期流行，且阳性率占比逐渐升高。测序分析提示，广州市H5亚型禽流感病毒为Clade 2.3.4.4.C分支H5N6高致病性病毒，并且持续进化和变异，特别是普遍出现与人源受体结合能力增强的突变，需加强监测。.
Keywords: Avian influenza virus; Epidemiology; Genetic evolution; Variation.