一株羊源A型产气荚膜梭菌的分子生物学诊断  

Molecular Biological Diagnosis of a Strain of Sheep-derived Clostridium Perfringens Type A

作　　者：王奎 WANG Kui(Dengzhou Animal Disease Prevention and Control Center,Dengzhou 474150,China)

机构地区：[1]邓州市动物疫病预防控制中心,河南邓州474150

出　　处：《乡村科技》2025年第1期104-107,共4页Rural Science and Technology

摘　　要：为查明河南省邓州市某羊场病死羊病因,尽快控制病情,采集病羊肝脏、肺脏和脾脏等器官组织样本进行细菌16S rDNAPCR扩增测序,并利用MEGA7.0软件构建系统进化树,PCR鉴定毒素基因分型。结果表明,在病料中分离鉴定出一株分离菌;对该分离菌株进行16S rDNA基因扩增,得到1400 bp阳性条带,与产气荚膜梭菌核苷酸同源性均在99.6%以上,进行核苷酸同源性比对,与产气荚膜梭菌参考菌株相似性在99.6%以上,表明为产气荚膜梭菌;PCR毒素基因分型得到324 bp阳性条带,符合α毒素基因特征,证实其为A型产气荚膜梭菌,命名为DENGZHOU202409;构建遗传进化树进行分析,发现DENGZHOU202409菌株与上海人源218002-301、印度豹源ATCC 13124处于同一小分支,亲缘关系较近。综上所述,DENGZHOU202409分离菌株为此次羊群感染致病的病原菌,为A型产气荚膜梭菌。In order to determine the cause of dead sheep in a sheep farm in Dengzhou City and control the spread of the disease as soon as possible,the samples of liver,lung,spleen and other or⁃gans and tissues of the diseased sheep were collected for bacterial 16S rDNA PCR amplification and sequencing.The phylogenetic tree was constructed using MEGA7.0 software,and the toxin genotyp⁃ing was identified by PCR.The results showed that a strain of isolated bacteria was isolated and iden⁃tified in the diseased material.The 16S rDNA gene of the isolated strain was amplified to obtain a 1400 bp positive band,and the nucleotide sequence homology with Clostridium perfringens was more than 99.6%.The nucleotide homology was compared with the reference strain of Clostridium perfrin⁃gens,and the similarity was more than 99.6%,indicating that it was Clostridium perfringens.A 324 bp positive band was obtained by PCR toxin genotyping,which was consistent with the characteristics of alpha toxin gene.It was confirmed to be Clostridium perfringens type A and named DENGZHOU202409.By constructing a genetic phylogenetic tree analysis,DENGZHOU202409 strain was in the same small branch with Shanghai human source 218002-301 and Indian leopard source ATCC 13124,and the genetic relationship was close.In summary,the isolated strain of DENGZHOU202409 was the pathogenic bacteria of this sheep infection.This experiment provided a clinical reference data for the diagnosis and treatment of Clostridium perfringens.

关 键 词：羊 产气荚膜梭菌 16S rDNA 毒素分型 

分 类 号：S826[农业科学—畜牧学] S855.3[农业科学—畜牧兽医]