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4株H9亚型禽流感病毒流行毒株的分离与鉴定
邓永,王嘉,侯力丹,刘丹,黄小洁,胡爱民,王乐元,翟天舒,毛娅卿,孔冬妮∗
0
(中国兽医药品监察所)
摘要:
2016~2018年分别从北京4个不同的禽场分离和收集了的4株血凝性分离物,经初步鉴定均为H9亚型禽流感病毒,分别定名为A/Chicken/Beijing/BJ/2016(H9)株(简称BJ161株)、A/Chicken/Beijing/BJ/2016(H9)株(简称BJ162株)、A/Chicken/Beijing/BJ/2017(H9)株(简称BJ17株)和A/Chicken/Beijing/BJ/2018(H9)株(简称BJ18株),将分离得到的4株H9亚型病毒流行株进行了传代及鉴定。结果显示,用针对H9亚型禽流感病毒HA基因的特异性引物对分离株进行RT-PCR扩增,分别获得特异性目的片段后进行测序分析,结果显示4株分离株经鉴定均为H9亚型禽流感病毒。4株分离株病毒含量在107.7EID50/0.1ml~108.3EID50/0.1ml之间,HA效价在8log2~9log2,4株分离株均与H9亚型禽流感阳性血清出现阳性反应,而与SPF鸡血清、ND阳性血清、EDS阳性血清、H5亚型及H7亚型禽流感阳性血清均呈现阴性反应;4株分离株经翅静脉接种6周龄的SPF鸡后未出现临床症状,攻毒后5天病毒分离均为阳性。试验结果证明,这4株分离株均为低毒力H9亚型禽流感病毒株。
关键词:  禽流感病毒,H9亚型,分离,纯化,鉴定
DOI:
投稿时间:2020-11-18修订日期:2021-02-02
基金项目:
Isolation and Identification of Four H9 Subtype Avian Influenza Virus Epidemic Strains
,,,,,,,翟天舒,毛娅卿,孔冬妮∗
(China Institute of veterinary Drug Control)
Abstract:
From 2016 to 2018, four hemagglutinating isolates were isolated and collected from four different poultry farms in Beijing. All of them were identified as H9 subtype avian influenza virus. They were named A/Chicken/Beijing/BJ/2016(H9) strain(BJ161 strain)、A/Chicken/Beijing/BJ/2016(H9) strain(BJ162 strain)、A/Chicken/Beijing/BJ/2017(H9) strain(BJ17 strain)和A/Chicken/Beijing/BJ/2018(H9) strain(BJ18 strain).Four H9 subtype viruses were isolated and identified. The results showed that the four isolates were identified as H9 subtype avian influenza virus by RT-PCR with specific primers for HA gene of H9 subtype avian influenza virus. The viral content of the four isolates was107.7EID50/0.1ml~108.3EID50/0.1ml and the HA titer was 8log2~9log2. All the four isolates showed positive reaction with H9 subtype avian influenza positive serum, but negative reaction with SPF chicken serum, ND positive serum, EDS positive serum, H5 subtype and H7 subtype avian influenza positive serum;the four isolates did not appear after being inoculated with SPF chicken serum, ND positive serum, EDS positive serum, H5 subtype avian influenza positive serum The clinical symptoms and virus isolation were positive 5 days after challenge. The results showed that the four isolates were all low virulence H9 subtype avian influenza virus strains.
Key words:  avian  influenza virus, H9 subtype, isolation, purification, identification

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