基于不同抗原的犬布鲁氏菌病免疫层析胶体金抗体试纸条的制备和比较  

Preparation and Comparison of Colloidal Gold Immunochromatographic Strips for Canine Brucellosis Based on Different Antigens

作　　者：郭晓涵 焉鑫 邵卫星[2] 殷德辉 闫昊 南文龙[2] 樊晓旭[2] 孙世雄 张培培 孙翔翔[2] 刘蒙达 张皓博 孙淑芳[2] 孙明军[2] Guo Xiaohan;Yan Xin;Shao Weixing;Yin Dehui;Yan Hao;Nan Wenlong;Fan Xiaoxu;Sun Shixiong;Zhang Peipei;Sun Xiangxiang;Liu Mengda;Zhang Haobo;Sun Shufang;Sun Mingjun(School of Public Health,Xuzhou Medical University,Xuzhou 221004,Jiangsu,China;China Animal Health and Epidemiology Center,Qingdao 266032,Shandong,China;Xinjang Animal Health Supervision Institution,Unumqi 830011,Xinjiang,China)

机构地区：[1]徐州医科大学公共卫生学院,江苏徐州221004 [2]中国动物卫生与流行病学中心,山东青岛266032 [3]新疆维吾尔自治区动物卫生监督所,新疆乌鲁木齐830011

出　　处：《中国动物检疫》2024年第5期90-95,共6页China Animal Health Inspection

基　　金：“十四五”国家重点研发计划项目(2021YFD1800402)。

摘　　要：为建立针对犬布鲁氏菌病的免疫层析胶体金快速检测方法,利用从犬种布鲁氏菌菌株提纯的膜蛋白和可溶性蛋白,原核表达纯化的外膜蛋白Omp31和BP26以及基于B细胞抗原肽的融合蛋白F14,分别制备了试纸条,然后采用犬布鲁氏菌病阳性和阴性血清,分别对上述试纸条进行敏感性和特异性比较。结果显示:Omp31抗原的敏感性最高,其次是F14和BP26,而全菌膜蛋白和全菌可溶性蛋白的敏感性较差;特异性上,使用由5种抗原制备的胶体金试纸条检测30份犬布鲁氏菌病阴性血清,结果均为阴性;5种试纸条与沙门氏菌、大肠杆菌(O157)、大肠杆菌(H7)、霍乱弧菌、副溶血弧菌、军团菌阳性血清均不发生交叉反应。另外,由F14和BP26制备的试纸条可识别羊布鲁氏菌病阳性血清,而由Omp31、全菌膜蛋白和全菌可溶性蛋白制备的试纸条均不能识别牛、羊布鲁氏菌病阳性血清。综上所述,Omp31、F14和BP26可作为制备犬布鲁氏菌病免疫层析胶体金检测卡的备选抗原。本研究为粗糙型布鲁氏菌引起的布鲁氏菌病的诊断提供了技术支撑。In order to develop a colloidal gold immunochromatographic method for rapid detection of canine brucellosis,strips were prepared respectively using membrane proteins and soluble proteins extracted from canine B.canis,outer membrane proteins of Omp31 and BP26 purified by prokaryotic expression system,as well as fusion protein of F14 based on B cell antigen peptides.The sensitivity and specificity of the strips were compared respectively using canine brucellosis-positive and-negative sera.The results showed that for sensitivity,Omp31 antigen was the highest,followed by F14 and BP26,but membrane proteins and soluble proteins were poor;for specificity,30canine brucellosis negative sera were tested by the colloidal gold strips prepared by five antigens,with negative results;there was no cross reaction to the positive sera of Salmonella,Escherichia coli(O157),Escherichia coli(H7),Vibrio cholerae,Vibrio parahaemolyticus and Legionella.In addition,the strips prepared based on F14 and BP26 protein could be used to recognize positive serum of sheep brucellosis,while those prepared based on Omp31protein,cell membrane protein and soluble protein failed for positive sera of both cattle and sheep brucellosis.In conclusion,Omp31,F14 and BP26 protein could be used as candidates to prepare immunochromatographic strips for canine brucellosis.The diagnosis of brucellosis caused by rough Brucella could be technically supported by the study.

关 键 词：犬布鲁氏菌病 犬种布鲁氏菌 外膜蛋白 免疫层析胶体金检测卡 

分 类 号：S852.614[农业科学—基础兽医学]