竞争ELISA和间接ELISA方法应用于牛布鲁氏菌病净化的研究

doi:10.11843/j.issn.0366-6964.2024.05.032

畜牧兽医学报 ›› 2024, Vol. 55 ›› Issue (5): 2146-2153.doi: 10.11843/j.issn.0366-6964.2024.05.032

竞争ELISA和间接ELISA方法应用于牛布鲁氏菌病净化的研究

赵灿奇^1,2, 冯宇¹, 吕浪¹, 李彦军³, 魏玉磊⁴, 丁家波¹, 陈祥⁵, 蒋卉^1*

1. 中国农业科学院北京畜牧兽医研究所, 北京 100193;
2. 云南省大理州动物疫病预防控制中心, 大理 671005;
3. 内蒙古阿拉善右旗动物疫病预防控制中心, 阿拉善右旗 737300;
4. 内蒙古阿拉善左旗宗别立镇综合保障和技术推广中心, 阿拉善左旗 753421;
5. 江苏省人兽共患病学重点实验室江苏高校动物重要疫病与人兽共患病防控协同创新中心, 扬州 225000

收稿日期:2023-07-11 出版日期:2024-05-23 发布日期:2024-05-27
通讯作者: 蒋卉，主要从事重要人兽共患病新型疫苗和诊断技术研究，E-mail:jianghui01@caas.cn
作者简介:赵灿奇(1984-)，男，白族，云南剑川人，兽医师，主要从事动物传染病流行病学研究，E-mail:zhaocanqi@163.com；冯宇(1989-)，男，山东兰山人，高级兽医师，主要从事布鲁氏菌新型诊断方法研究。E-mail:13051530440@163.com。赵灿奇和冯宇为同等贡献作者
基金资助:
少数民族专业人才特殊培养计划项目；江苏省人兽共患病学重点实验室资助项目(R2210)

Study on Purification of Bovine Brucellosis by Competitive ELISA and Indirect ELISA

ZHAO Canqi^1,2, FENG Yu¹, LÜ Lang¹, LI Yanjun³, WEI Yulei⁴, DING Jiabo¹, CHEN Xiang⁵, JIANG Hui^1*

1. Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China;
2. Animal Disease Prevention and Control Center of Dali Prefecture, Yunnan Province, Dali 671005, China;
3. Animal Disease Prevention and Control Center of Alxa Youqi, Inner Mongolia, Alxa Youqi 737300, China;
4. Comprehensive Security and Technology Promotion Center in Zongbieli Town, Alxa Zuoqi, Inner Mongolia, Alxa Zuoqi 753421, China;
5. Jiangsu University's Collaborative Innovation Center for the Prevention and Control of Important Animal Diseases and Zoonosis, Jiangsu Key Laboratory of Zoonosis, Yangzhou 225000, China

Received:2023-07-11 Online:2024-05-23 Published:2024-05-27

摘要/Abstract

摘要： 旨在评价竞争ELISA方法和间接ELISA方法联合使用在布鲁氏菌病(以下简称“布病”)净化中的效果，为布病防控提供技术支撑。采用本实验室开发的动物布鲁氏菌病竞争ELSIA(cELISA)抗体检测试剂盒、牛布鲁氏菌病间接ELISA(iELISA)抗体检测试剂盒及微量法补体结合试验(mCFT)，对西北某牛场3 271份牛血清进行检测。本研究采用cELISA初筛、iELISA确诊的净化策略，对检测阳性牛进行淘汰，可疑牛和阴性牛在完全消毒后隔离饲养，并在前一次检测后每隔1个月重新对群体采样，进行多次连续的“检—淘”策略，在群体的个体阳性率低于2%或全部转阴后使用微量补体结合试验(mCFT)进行验证。并在群体全部转阴后继续检测2个月后确定净化结果。结果显示，首次检测阳性率35.36%的感染群体实施本净化策略后，在第1个月阳性率下降至25.41%，第2个月下降至7.16%，第3个月下降为1.86%，到第4个月则实现了布病阳性群体的全面转阴，mCFT验证个体阴性率100%。此后持续检测2个月，个体阳性率均为0，至此实现了感染群体的布病净化，使得群体内近一半的牛免于扑杀，大大减少了经济损失。综上发现，将cELISA用于布病初筛，iELISA用于布病确诊的联合使用，经多次连续检测并结合常规的隔离消毒措施，可以在短时间内实现对于布病感染群体的全面净化。

关键词: 布鲁氏菌病, 竞争酶联免疫吸附试验, 间接酶联吸附试验, 微量补体结合试验, 初筛, 确诊

Abstract: The aim of this study was to evaluate the efficacy of the combination of competitive ELISA and indirect ELISA methods in the purification of brucellosis, hoping to provide technical support for the prevention and control of brucellosis. A total of 3 271 bovine serum samples from a cattle farm in the northwest of China were tested using the competitive ELISA (cELISA) antibody detection kit, the indirect ELISA (iELISA) antibody detection kit, and the micro-complement fixation test (mCFT) which developed by the writers team. This study adopted a purification strategy of cELISA for preliminary screening, iELISA for confirm diagnosis. The positive cattle were eliminating, the suspect cattle and negative cattle were respectively quarantined after disinfecting completely. After the previous detection, the group was sampled every month for multiple “Detection-Elimination” cycles. When the individual positive rate was below 2% or all negative, the results were verified using a micro-complement fixation test (mCFT). The test were carried on for 2 months after all the bovine turned negative to ensure the successful of the purification. The results showed that, after the implementation of this purification strategy, the positive rate of the infected group with a positive rate of 35.36% decreased to 25.41% in the first month, 7.16% in the second month, and 1.86% in the third month. By the fourth month, the group had achieved a comprehensive negative conversion, and the negative rate by mCFT verification was 100%. The detection continued for 2 months, and the individual positive rate was 0. So far, the brucellosis purification of the infected group was realized, making nearly half of the cattle in the group free from culling, greatly reducing the economic loss. In conclusion, cELISA is suitable for the preliminary screening, while iELISA is suitable for confirmed diagnosis of brucellosis. The combination of two methods, through multiple consecutive tests, can achieve a comprehensive purification of brucellosis infected within a relatively short period of time.

Key words: brucellosis, competitive ELISA, indirect ELISA, micro-complement fixation test, preliminary screening, confirmed diagnose

中图分类号:

S852.614

赵灿奇, 冯宇, 吕浪, 李彦军, 魏玉磊, 丁家波, 陈祥, 蒋卉. 竞争ELISA和间接ELISA方法应用于牛布鲁氏菌病净化的研究[J]. 畜牧兽医学报, 2024, 55(5): 2146-2153.

ZHAO Canqi, FENG Yu, LÜ Lang, LI Yanjun, WEI Yulei, DING Jiabo, CHEN Xiang, JIANG Hui. Study on Purification of Bovine Brucellosis by Competitive ELISA and Indirect ELISA[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2146-2153.

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竞争ELISA和间接ELISA方法应用于牛布鲁氏菌病净化的研究

Study on Purification of Bovine Brucellosis by Competitive ELISA and Indirect ELISA

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