鸭多杀性巴氏杆菌抗体检测胶体金试纸条的研制及应用  

Development and Application of Colloidal Gold Test Strip for Detecting Antibodies to Pasteurella multocida in Duck Serum

作　　者：程龙飞[1] 陈红梅[1] 傅光华[1] 江南松 傅秋玲[1] 万春和[1] 黄瑜[1] 刘荣昌[1] 施少华[1] CHENG Long-fei;CHEN Hong-mei;FU Guang-hua;JIANG Nan-song;FU Qiu-ling;WAN Chun-he;HUANG Yu;LIU Ron-chang;SHI Shao-hua(Fujian Provincial Key Laboratory for Avian Diseases Control and Prevention,Fujian Animal Diseases Control Technology Center,Institute of Animal Husbandry and Veterinary Medicine,Fujian Academy of Agricultural Sciences,Fuzhou,Fujian,350013,China)

机构地区：[1]福建省农业科学院畜牧兽医研究所/福建省禽病防治重点实验室/福建省畜禽疫病防治工程技术研究中心,福建福州350013

出　　处：《动物医学进展》2023年第6期65-69,共5页Progress In Veterinary Medicine

基　　金：国家水禽产业技术体系项目(CARS-42);福建省科技厅农业科技重大专项项目(2019NZ09008);公益类科研院所专项项目(2020R1026005)。

摘　　要：为建立快速检测鸭血清多杀性巴氏杆菌抗体的方法,将兔抗鸭IgG标记胶体金,喷涂金标垫,用浓度为0.2 mg/mL的荚膜和0.2 mg/mL羊抗兔IgG抗体分别作为检测线和质控线,制备了检测鸭多杀性巴氏杆菌抗体的胶体金试纸条(D-CGTS)。特异性试验显示,仅鸭抗多杀性巴氏杆菌血清检测为阳性,阴性血清、鸭抗大肠埃希氏菌、鸭抗鸭疫里默氏菌、鸭抗沙门氏菌和鸭抗金黄色葡萄球菌血清的检测结果均为阴性,表明D-CGTS的特异性良好。敏感性试验结果显示,对效价1∶16(琼脂扩散试验)的鸭抗多杀性巴氏杆菌血清1∶320倍稀释后采用制备的试纸条检测结果仍为阳性,表明D-CGTS的敏感性较高。重复性试验结果显示,同批次的不同试纸条和不同批次的试纸条,检测阴性血清和鸭抗多杀性巴氏杆菌血清的结果相同,表明D-CGTS的重复性好。对92份鸭血清采用D-CGTS和ELISA进行检测,结果显示,阳性符合率为94.03%,阴性符合率为100%,总符合率为95.65%。结果表明,制备的D-CGTS操作简单,具有较强的特异性、较高的敏感性和较好的重复性,适合在兽医临床推广应用。In order to establish a method for rapid detection of antibodies to Pasteurella multocida in duck serum,colloidal gold-labeled rabbit anti-duck IgG was used in this study to prepare gold-labeled pads.Capsule(0.2 mg/mL)and goat anti-rabbit IgG(0.2 mg/mL)were coated as detection line and quality line respectively,then were assembled into colloidal gold test strips(D-CGTS)after optimizing conditions.The method was used to detect the negative sera,the positive sera of Pasteurella multocida,Escherichia coli,Riemerella anatipestifer,Salmonella and Staphylococcus aureus of ducks.The results showed that only Pasteurella multocida positive sera were positive,and all of the others were negative,indicating the high specificity of D-CGTS.The positive serum of Pasteurella multocida with the agar diffusion(AGP)titer of 1∶16 was detected by D-CGTS after dilution.The sensitivity results showed that the 1∶320 times dilution could still detect positive,indicating that the sensitivity of D-CGTS was high.There was no difference between different test strips in the same batch and different batches in detection of Pasteurella multocida positive serum and negative serum,indicating that the method has good repeatability.92 duck sera were detected by D-CGTS and ELISA.The results showed that the positive coincidence rate was 94.03%,the negative coincidence rate was 100%,and the total coincidence rate was 95.65%.In conclusion,the D-CGTS developed in this study shows the advantages of simple operation,strong specificity,good repeatability,and high sensitivity in detection of antibodies to Pasteurella multocida in duck serum,which is suitable for clinical application.

关 键 词：鸭 多杀性巴氏杆菌 胶体金试纸条 荚膜 

分 类 号：S852.612[农业科学—基础兽医学] S858.33[农业科学—兽医学]