一起羊黑疫的病原分离诊断及毒株产毒性能分析  

Pathogen isolation and PCR Diagnosis of Black Disease in Sheep

作　　者：李淑萍 张西云[1] 胡国元[1] 贾苗 叶明俊 河生德[3] 李长云[3] 祁果 李生庆[1] LI Shu-ping(Academy of Animal Science and Veterinary Medicine of Qinghai University,Xining 810016)

机构地区：[1]青海大学畜牧兽医科学院,西宁810016 [2]青海大学农牧学院,西宁810013 [3]青海省海晏县畜牧兽医站,海晏812000

出　　处：《青海畜牧兽医杂志》2019年第6期27-31,共5页Chinese Qinghai Journal of Animal and Veterinary Sciences

基　　金：家畜主要疫病诊断与防控技术研究示范(2017-NK-110)

摘　　要：利用厌氧菌实验室常规检测方法从一例送检病死羊肾脏培养物中分离到一株梭杆菌。通过形态学观察、动物实验、血清中和及胰酶破坏试验,确定分离病原菌为B型诺维氏梭菌。用自行设计的诺维氏梭菌α毒素特异性引物对纯培养物进行PCR扩增检测,PCR扩增得到大小为530bp的目的条带,与常规诊断结果一致;对分离的野生毒株的产毒能力进行了测定分析,野毒分离株在自制的VF厌气肝汤中的产毒能力达4000 MLD/mL,证明该菌株具备生产羊黑疫疫苗用株的潜力。A strain of Clostridium was isolated from a kidney culture of a dead sheep by an routine experiment of anaerobic laboratory. Through morphological observation, animal experiments, serum neutralization and trypsin destruction test, it was determined that the isolated pathogen was Clostridium Novyi type B, and PCR was used to detect the pure culture with the self-designed Clostridium Novyi a toxin specific primer. PCR amplification can get the target band of 530 bp, which was consistent with the routine diagnosis results. The toxigenic ability of the isolated wild strain was determined and analyzed. The produce toxigenic quantity of the wild stains in the homemade VF anaerobic liver soup was 4000 MLD/mL. The result showed that this strain have the potential as a black sheep disease vaccine strain in the future.

关 键 词：羊黑疫 病原分离 PCR诊断 

分 类 号：S852.61[农业科学—基础兽医学]