商品疫苗对我国h9.4.2.5分支H9N2亚型禽流感分离株的免疫保护  

The Protective Efficacy of Commercial Vaccines Against H9N2 Avian Influenza Virus of Branch h9.4.2.5 Isolated in China

作　　者：麻琦 和新文 王燕 刘艳晶 潘舒心 侯玉杰[1] 施建忠[1,3] 邓国华 包红梅[1,3] 刘景利 郭兴福[2] 毛胜刚 胡井雷 路通 杨帆 田国彬 曾显营 陈化兰 MA Qi;HE XinWen;WANG Yan;LIU YanJing;PAN ShuXin;HOU YuJie;SHI JianZhong;DENG GuoHua;BAO HongMei;LIU JingLi;GUO XingFu;MAO ShengGang;HU JingLei;LU Tong;YANG Fan;TIAN GuoBin;ZENG XianYing;CHEN HuaLan(National Poultry Laboratory Animal Resource Center/State Key Lab for Animal Disease Control and Prevention/Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150069;Harbin Weike Biotechnology Company Limited,Harbin 150069;Key Laboratory for Prevention and Control of Avian Influenza and Other Major Poultry Diseases,Ministry of Agriculture and Rural Affairs,Harbin 150069)

机构地区：[1]中国农业科学院哈尔滨兽医研究所/动物疫病防控全国重点实验室/国家禽类实验动物资源库,哈尔滨150069 [2]哈尔滨维科生物技术有限公司,哈尔滨150069 [3]农业农村部禽流感等家禽重大疾病防控重点实验室,哈尔滨150069

出　　处：《中国农业科学》2023年第15期3040-3050,共11页Scientia Agricultura Sinica

基　　金：国家重点研发计划(2021YFD1800200);现代农业产业技术体系(CARS-41)。

摘　　要：【目的】我国批准使用的H9亚型禽流感(avian influenza,AI)商品化灭活疫苗种类繁多,其免疫效果和选用受到养殖者的广泛关注。通过评估不同商品疫苗对我国近期H9N2亚型AIV的免疫保护效果,以期为H9亚型AIV的免疫防控提供科学参考。【方法】依据国家兽药基础数据库疫苗批签发数据,从在售的40种H9亚型AI商品疫苗中选择批签发数量较多的4种疫苗(A—D疫苗),进行免疫攻毒试验。4株h9.4.2.5分支的H9N2亚型分离株CK/XJ/S1204/2015、DK/JX/S4512/2017、CK/YN/S1666/2020和CK/NX/S4590/2020为攻毒毒株,分别测定4株病毒的鸡胚半数感染量(EID_(50))、鸡半数感染量(CID50)和细胞半数感染量(TCID50),以确定动物试验的攻毒剂量和细胞试验的感染剂量。每种疫苗以产品推荐剂量免疫3周龄SPF鸡各40只,同时设同日龄SPF鸡40只接种PBS作为对照组;免疫3周时,采集所有试验鸡血清,测定血凝抑制抗体(HI)和中和抗体(NT)滴度;同时将每种商品疫苗接种40只SPF鸡进行随机分组,每组10只,连同10只对照组鸡,以10 CID50的剂量鼻腔感染H9N2亚型分离株,分别进行商品疫苗对4株病毒的攻毒保护试验。采集攻毒后3、5 d喉头及泄殖腔棉拭子样品,接种10日龄鸡胚检测排毒情况,统计疫苗保护率,比较疫苗对H9N2亚型分离株的免疫保护效果。【结果】4株H9N2亚型分离株的CID50依次为10^(3.5)、10^(2.5)、10^(2.5)和10^(3.5)EID_(50)/0.1 mL。商品疫苗接种后3周,各免疫组SPF鸡血清中针对商品化H9亚型HI试验抗原(CK/SH/10/2001)的HI抗体均在9.4 log_(2)—11 log_(2)之间,但针对攻毒株的HI抗体平均效价在4.6 log_(2)—10.8 log_(2)之间,不同疫苗免疫组间存在较大差异,最大差异为64倍,各免疫组NT抗体平均效价在6.7 log_(2)—12.2 log_(2)之间,最大差异为32倍,对照组鸡的HI抗体和NT抗体均为阴性。以滴鼻方式感染不同H9病毒后,4种疫苗的免疫保护效果存在较大差别,在攻击CK/XJ/S【Objective】There are many commercial inactivated vaccines against the H9 subtype avian influenza approved for use in China,and their protection efficacy and selection have been widely concerned by farmers.In this study,the protection efficacy of main commercial vaccines against H9N2 viruses recently isolated in the field were evaluated,so as to provide the research reference for guiding the prevention and control of H9 subtype AI by immunization in China.【Method】According to the vaccine batch release of the National Veterinary Drug Basic Information Database in China,4 commercial vaccines(labeled as A-D)with large batches were selected from 40 kinds of sold H9 commercial vaccines.The four H9N2 subtype AIV,including CK/XJ/S1204/2015,DK/JX/S4512/2017,CK/YN/S1666/2020 and CK/NX/S4590/2020,belonging to h9.4.2.5 branch and isolated at different times and places,were used to evaluate the protection efficacy of selected commercial H9 subtype AI vaccines.The50%chicken embryo infection(EID_(50)),50%chicken infection(CID_(50)),and 50%cell infection(TCID_(50))of the four viruses were tested to determine the challenge dose of the animal experiment and the infection dose of the cell test.403-week-old SPF chickens were injected intramuscularly with 4 kinds of inactivated vaccine,and the groups of 10 similar SPF chickens were inoculated with PBS as control group.3 weeks post vaccination(p.v.),the serum of all experimental chickens were collected before challenge,the hemagglutination inhibition(HI)and neutralization(NT)antibody titers were detected by HI test and neutralization test;the 40 immunized chickens of each vaccine were randomly divided into four groups,and 10 chickens of each vaccine and 10 control chickens were infected intranasally with the 10 CID_(50)of each H9N2 virus.Oropharyngeal and cloacal swabs were collected on days 3 and 5 post challenge(p.c.)for virus shedding detection and calculation of the protective rate of each vaccine.【Result】The CID_(50)of the four strains were 10^(3.5)EID_(50)/0.1 mL,10^

关 键 词：H9N2亚型 AIV 商品疫苗 免疫效果 禽流感 

分 类 号：S858.31[农业科学—临床兽医学]