布鲁氏菌病M5、S2疫苗免疫血清半抗原-琼脂扩散试验方法检测评价  

Evaluation of sera immunized with Brucella M5 and S2 vaccines with native hapten-agar gel immunodiffusion tests

作　　者：刘丽娅[1] 剡文亮[2] 张子荣[3] 宋洁 郑莹 马晓菁[1] 叶锋[1] 谷文喜[1] 易新萍[1] 李岩[2] LIU Li-ya;YAN Wen-liang;ZHANG Zi-rong;SONG Jie;ZHENG Ying;MA Xiao-jing;YE Feng;GU Wen-xi;YI Xin-ping;LI Yan(Xinjiang Key Laboratory of Animal Infectious Diseases,Institute of Veterinary Medicine,Xinjang Academy of Animal Science,Urumqi 830011,China;The General Station of Animal Husbandry and Veterinary of Xinjiang Production and Construction Corps,Urumqi 830021,China;Xinjiang Production and Construction Corps Animal Husbandry and Veterinary Work Station of the Sixth Division,Wujiaqu 831304,China;Xinjiang Tianrun Dairy Limited Company,Urumqi 830000,China)

机构地区：[1]新疆畜牧科学院兽医研究所,新疆动物疫病研究重点实验室,乌鲁木齐830011 [2]新疆生产建设兵团畜牧兽医工作总站,乌鲁木齐830021 [3]新疆生产建设兵团第六师畜牧兽医工作站,五家渠831304 [4]新疆天润乳业股份有限公司,乌鲁木齐830000

出　　处：《中国人兽共患病学报》2023年第8期766-771,共6页Chinese Journal of Zoonoses

基　　金：新疆生产建设兵团重点领域科技攻关计划项目(No.2020AB015)。

摘　　要：目的应用NH-AGID方法对布鲁氏菌M5、S2疫苗株免疫血清进行检测,评价该方法适用性。方法选取3~5月龄羔羊74只,随机分为3组。分别以S2株1.0×1010 CFU、5.0×109 CFU灌服和皮下注射接种羔羊各25只,M5株以1.0×10^(9) CFU皮下注射接种羔羊24只。免疫后7 d~150 d采集羊全血,分离血清,经RBT初筛和SAT确诊免疫抗体阳性血清。评估NH-AGID方法对M5和S2株疫苗免疫抗体与感染抗体的鉴别诊断特异性。结果NH-AGID方法对M5株疫苗免疫阳性血清的LPS抗体检出率为100%,NH抗体检出率8.3%~29.2%,鉴别诊断特异性为82.8%。对S2株疫苗口服组免疫阳性血清的LPS抗体检出率为31.2%~66.7%,NH抗体检出率为0%;皮下注射组免疫阳性血清的LPS抗体检出率为45.4%~100%,NH抗体检出率为4.5%~20%。对S2株疫苗免疫血清的鉴别诊断特异性为96.5%。结论NH-AGID方法不完全适用于羊用布鲁氏菌疫苗M5株和S2株的鉴别诊断,该方法的鉴别诊断特异性与疫苗毒力、免疫剂量和途径等因素相关,应进一步研究明确该方法的适用范围及条件,以期合理应用于布病防控诊断工作中。The native hapten-agar gel immunodiffusion test was used to detect sera immunized with Brucella M5 and S2 vaccine strains,to evaluate the applicability of this method.A total of 74 sheep 3-5 months old were randomly divided into three groups;25 sheep were inoculated with the S2 vaccine strain,1.0×1010 CFU filling and 5.0×10^(9)CFU subcutaneous injection;24 sheep were inoculated subcutaneously with the M5 vaccine strain,1.0×109 CFU.Sheep blood was collected from 7 to 150 days after immunization,the serum was separated,and RBT preliminary screening and the SAT method were used to detect immune antibody positive serum.The NH-AGID method was used to evaluate the specificity of differential diagnosis between the M5 and S2 vaccine immune antibody and infection antibody.The detection rate of LPS antibody in M5 vaccine positive serum with the NH-AGID method was 100%,the detection rate of NH antibody was 8.3%-29.2%,and the specificity of differential diagnosis was 82.8%.The detection rates of LPS antibody and NH antibody in immune positive sera in the S2 vaccine strain oral group were 31.2%-66.7%and 0%respectively.The detection rates of NH antibody and LPS antibody in the subcutaneous injection group were 4.5%-20%and 45.4%-100%,respectively.The specificity of differential diagnosis of S2 vaccine strain immune serum was 96.5%.The NH-AGID method is not completely applicable to the differential diagnosis of the immune antibodies and infection antibodies of S2 and M5 strains of Brucella ovis vaccine in China.The specificity of the differential diagnosis of this method is related to many factors such as vaccine virulence,immune dose and pathway.Further research should be performed to clarify the scope and conditions of application of this method,to enable its application to the prevention and diagnosis of brucellosis.

关 键 词：布鲁氏菌病 半抗原-琼脂扩散试验 M5株 S2株 鉴别诊断 

分 类 号：S855.1[农业科学—临床兽医学]