奶牛源化脓隐秘杆菌的分离鉴定及其毒力基因的检测

›› 2014, Vol. 41 ›› Issue (11): 68-72.

奶牛源化脓隐秘杆菌的分离鉴定及其毒力基因的检测

李伟杰¹, 魏财文¹, 刘燕², 岂晓鑫¹, 田野¹, 蒋桃珍¹

1. 中国兽医药品监察所, 北京 100081;
2. 北京动物园, 北京 100081

收稿日期:2014-06-16 出版日期:2014-11-20 发布日期:2014-12-06
通讯作者: 蒋桃珍 E-mail:jiangtaozhen@ivdc.gov.cn
作者简介:李伟杰(1979-),男,山东人,硕士,助理研究员,研究方向:兽医微生物学。
基金资助:
国家微生物资源平台(NIMR-5)。

Isolation, Identification of Trueperella pyogenes from Dairy Cow and Detection of Major Virulence Genes of the Isolate

LI Wei-jie¹, WEI Cai-wen¹, LIU Yan², QI Xiao-xin¹, TIAN Ye¹, JIANG Tao-zhen¹

1. China Institute of Veterinary Drug Control, Beijing 100081, China;
2. Beijing Zoo, Beijing 100081, China

Received:2014-06-16 Online:2014-11-20 Published:2014-12-06

摘要/Abstract

摘要： 本研究旨在分离鉴定来自北京某奶牛场死亡奶牛肺脏的1株疑似病原菌CVCC 3982,并测定其致病性。通过分离培养获得疑似病原菌,采用Biolog鉴定系统和16S rDNA序列分析对其进行了鉴定,人工接种CD-1小鼠测定了其致病性,合成引物对其主要毒力基因进行了检测。结果显示,该疑似病原菌为革兰氏阳性杆菌,β溶血,Biolog鉴定结果显示其为化脓隐秘杆菌,其16S rDNA序列与化脓隐秘杆菌模式菌株NCTC 5224的同源性达100%,系统发育分析显示其与化脓隐秘杆菌处于同一分支。腹腔注射该菌可致小鼠死亡。分离菌株基因组中含有溶血素(PLO)基因,神经氨酸酶H(NanH)基因,神经氨酸酶P(NanP)基因,菌毛基因(fimA、fimC和fimE),但缺失胶原结合蛋白(CbpA)基因和菌毛fimG基因。结果表明该分离菌株为化脓隐秘杆菌且具有致病性。

关键词: 奶牛; 化脓隐秘杆菌; Biolog; 16S rDNA; 毒力基因

Abstract: The purpose of this study was to identify the strain CVCC 3982 isolated from a death dairy cow's lung in Beijing and determine its pathogenicity. The suspected pathogen was identified by means of Biolog system and 16S rDNA sequence analysis. Its pathogenicity was determined by artificial inoculation in CD-1 mice, synthetic primers were used to examine the main virulence genes. The results showed that the suspected pathogen was gram positive bacillus, β hemolysis. The identification result of Biolog was Trueperella pyogenes. The homology of 16S rDNA gene sequence with the Trueperella pyogenes type strain NCTC 5224 reached 100%. By the phylogenetic analysis, the isolate was in the same branch with Trueperella pyogenes. The isolate contained hemolysin (plo) gene, neuraminidase H (nanH) gene, neuraminidase P (nanP) gene, fimbriae gene (fimA, fimC, fimE), but was lack of collagen-binding protein (cbpA) gene and fimG gene. Infection of suspected bacteria resulted in mortality to mice by intraperitoneal injection. The results indicated that the isolate was a virulent Trueperella pyogenes.

Key words: dairy cow; Trueperella pyogenes; Biolog; 16S rDNA; virulence genes

中图分类号:

S852.61

李伟杰, 魏财文, 刘燕, 岂晓鑫, 田野, 蒋桃珍. 奶牛源化脓隐秘杆菌的分离鉴定及其毒力基因的检测[J]. , 2014, 41(11): 68-72.

LI Wei-jie, WEI Cai-wen, LIU Yan, QI Xiao-xin, TIAN Ye, JIANG Tao-zhen. Isolation, Identification of Trueperella pyogenes from Dairy Cow and Detection of Major Virulence Genes of the Isolate[J]. , 2014, 41(11): 68-72.

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