摘要
将羊肺脏中分离鉴定后的肺炎克雷伯菌制成菌体免疫抗原,免疫健康家兔后获得兔抗羊源肺炎克雷伯菌多克隆抗体。依据此多克隆抗体建立检测肺炎克雷伯菌的间接免疫荧光检测方法,并对检测条件进行了筛选和优化。结果表明,多克隆抗体稀释度为1∶80,FITC标记羊抗兔IgG稀释度为1∶400,孵育时间为30 min时,羊源肺炎克雷伯菌的特异性黄绿色荧光最清晰,非特异性荧光最弱。并通过临床检测证明该方法具有简便、特异性高的优点,为羊源肺炎克雷伯菌的流行病学、血清学调查和临床诊断奠定基础。
Klebsiella pneumoniae(K.pneumoniae)can cause disease in humans and a variety of animals under certain conditions,but there is no relevant research on the detection method of Klebsiella pneumoniae of sheep origin.In this study,a polyclonal antibody against Klebsiella pneumoniae was obtained by immunizing healthy rabbits with Klebsiella pneumoniae isolated from lungs of sheep.An indirect immunofluorescence assay for detecting Klebsiella pneumoniae was established based on this polyclonal antibody,and the assay conditions were optimized.The results showed that under the conditions of 80 times dilution of polyclonal antibody,400 times dilution of FITC-labeled goat anti-rabbit IgG and incubation for 30 min,the specific yellow-green fluorescence of Klebsiella pneumoniae was the clearest and the non-specific fluorescence was the weakest.Furthermore,the method was proved to be simple and specific by clinical tests,which laid a foundation for the epidemiological and serological investigation and clinical diagnosis of Klebsiella pneu moniae.
作者
陈瑞昶
牛祥云
黄金
苗永强
胡莉萍
朱瑞良
CHEN Rui-chang;NIU Xiang-yun;HUANG Jin;MIAO Yong-qiang;HU Li-ping;ZHU Rui-liang(Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention,College of Animal Science and Technology,Shandong Agricultural University,Tai'an,Shandong 271018,China;Shandong Animal Disease Prevention and Control Center,Jinan 250022,China)
出处
《中国兽医学报》
CAS
CSCD
北大核心
2020年第12期2342-2347,共6页
Chinese Journal of Veterinary Science
基金
国家“十三五”重点研发计划基金资助项目(2016YFD0500905)
山东省现代农业产业技术体系羊产业创新团队建设基金资助项目(SDAIT-10-06)。
关键词
羊源
肺炎克雷伯菌
多克隆抗体
间接免疫荧光
Sheep source
Klebsiella pneumoniae
polyclonal antibody
indirect immunofluorescence