羊源D型多杀性巴氏杆菌的分离鉴定及毒力基因检测
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摘要
为了确定石河子地区规模化羊场出现呼吸道症状死亡羊的细菌性病原,采用常规细菌分离鉴定方法、细菌16SrRNA序列分析以及多杀性巴氏杆菌特异性基因kmt,从病变肺组织分离鉴定细菌,利用5个荚膜血清型特异性基因确定其血清型,扩增分离株的16个毒力相关基因,分析分离菌致病性和对常用抗菌药物的耐药性。结果表明,从病羊的病变肺组织中分离鉴定到一株血清D型多杀性巴氏杆菌,具有较强的致病性;携带8个毒力相关基因,其片段序列与NCBI上己公布的多杀性巴氏杆菌参考株同源性在99%以上;分离株对青霉素、林可霉素、庆大霉素、复方新诺明耐药,对其他23种药物敏感。
In order to determine the cause of the sheep died of respiratory infection from a sheep farm in Shihezi,bacterial pathogens were isolated and identified by conventional methods and molecular biological methods based on 16 SrRNA special sequence and Pm specific gene kmt sequence analysis.Serotype of the isolate was determined with the primers from 5capsular serotype-specific genes,16virulence-associated genes were detected,pathogenicity and drug sensitivity of the isolate were analysed.The results showed that the isolate was a serotype D Pm with strong pathogenicity,which carried 8virulence-associated genes and the amplified fragment homology of the corresponding sequence in NCBI is more than 99%.The isolate was resistant to penicillin,lincomycin,gentamicin,cotrimoxazole.
In order to determine the cause of the sheep died of respiratory infection from a sheep farm in Shihezi,bacterial pathogens were isolated and identified by conventional methods and molecular biological methods based on 16 SrRNA special sequence and Pm specific gene kmt sequence analysis.Serotype of the isolate was determined with the primers from 5capsular serotype-specific genes,16virulence-associated genes were detected,pathogenicity and drug sensitivity of the isolate were analysed.The results showed that the isolate was a serotype D Pm with strong pathogenicity,which carried 8virulence-associated genes and the amplified fragment homology of the corresponding sequence in NCBI is more than 99%.The isolate was resistant to penicillin,lincomycin,gentamicin,cotrimoxazole.
引文
[1] Marina H,Harper M,John M,et al.Development of a rapid multiplex PCR assay to genotype Pasteurella multocidastrains by use of the lipopolysaccharide outer core biosynthesis locus[J].J Clin Microbiol,2015(53),477-485.
[2]孙宝金.羊巴氏杆菌病的诊断与防治[J].畜牧兽医科技信息,2014(1):42-43.
[3] Townsend K M,Boyce J D,Chung J Y,et al.Genetic organization of Pasteurella multocida cap loci and development of a multiplex capsular PCR typing system[J].J Clin Microbiol,2001,39(3):924-929.
[4] Watt J M,Swiatlo E,Wade M M,et al.Regulation of capsule biosynthesis in serotype A strains of Pasteurella multocida[J].FEMS Microbiol Lett,2003,225(1):9-14.
[5]马文戈,于力.牛源荚膜血清A型多杀性巴氏杆菌的分离鉴定[J].中国预防兽医学报,2008(10):747-750,754.
[6]杨楠,吴同垒,王洪彬,等.鹅源多杀性巴氏杆菌的分离鉴定与药物敏感性分析[J].中国家禽,2017,39(22):55-58.
[7]阿依江·卡那提拜克,姚刚,王世民.新疆某羊场巴氏杆菌的分离、鉴定与药敏试验[J].中国草食动物科学,2017,37(5):41-44.
[8]刘顺磊,何延华,王熙楚,等.新疆石河子规模化牛场多杀性巴氏杆菌的分离鉴定及药敏试验[J].动物医学进展,2016,37(6):40-44.
[9]肖淦文,陈颖钰,彭清洁,等.牛支原体、巴氏杆菌A型和化脓隐秘杆菌多重PCR快速检测方法的建立[J].中国奶牛,2012(21):4-9.
[10]潘婷婷,冯思源,杜慧慧,等.牛源A型多杀性巴氏杆菌强毒株CQ2和弱毒株CQ6毒力相关基因分析[J].中国奶牛,2015(Z3):22-24.
[11] Khamesipour F,Momtaz H,Mamoreh M A.Occurrence of virulence factors and antimicrobial resistance in Pasteurella multocida strains isolated from slaughter cattle in lran[J].Front Microbiol,2014(5):536-536.
[12]胡继红,高振翔,尹铭芳.美国NCCLS2002年版抗生素药敏试验操作标准更新内容[J].中华检验医学杂志,2002(6):46-48.
[13] Sarangi L N,Priyadarshini A,Kumar S,et al.Virulence genotyping of Pasteurella multocida lsolated from multiple hosts from India[J].Sci World J,2014(2014):814109-814109.
[14]王羽,董文龙,王巍,等.牛源多杀性巴氏杆菌荚膜血清型及毒力基因检测[J].中国兽医杂志,2017,53(9):16-19.
[15]李伟杰,赵耘,杜昕波,等.产毒素多杀性巴氏杆菌的鉴定[J].中国畜牧兽医,2010,37(1):149-151.
[2]孙宝金.羊巴氏杆菌病的诊断与防治[J].畜牧兽医科技信息,2014(1):42-43.
[3] Townsend K M,Boyce J D,Chung J Y,et al.Genetic organization of Pasteurella multocida cap loci and development of a multiplex capsular PCR typing system[J].J Clin Microbiol,2001,39(3):924-929.
[4] Watt J M,Swiatlo E,Wade M M,et al.Regulation of capsule biosynthesis in serotype A strains of Pasteurella multocida[J].FEMS Microbiol Lett,2003,225(1):9-14.
[5]马文戈,于力.牛源荚膜血清A型多杀性巴氏杆菌的分离鉴定[J].中国预防兽医学报,2008(10):747-750,754.
[6]杨楠,吴同垒,王洪彬,等.鹅源多杀性巴氏杆菌的分离鉴定与药物敏感性分析[J].中国家禽,2017,39(22):55-58.
[7]阿依江·卡那提拜克,姚刚,王世民.新疆某羊场巴氏杆菌的分离、鉴定与药敏试验[J].中国草食动物科学,2017,37(5):41-44.
[8]刘顺磊,何延华,王熙楚,等.新疆石河子规模化牛场多杀性巴氏杆菌的分离鉴定及药敏试验[J].动物医学进展,2016,37(6):40-44.
[9]肖淦文,陈颖钰,彭清洁,等.牛支原体、巴氏杆菌A型和化脓隐秘杆菌多重PCR快速检测方法的建立[J].中国奶牛,2012(21):4-9.
[10]潘婷婷,冯思源,杜慧慧,等.牛源A型多杀性巴氏杆菌强毒株CQ2和弱毒株CQ6毒力相关基因分析[J].中国奶牛,2015(Z3):22-24.
[11] Khamesipour F,Momtaz H,Mamoreh M A.Occurrence of virulence factors and antimicrobial resistance in Pasteurella multocida strains isolated from slaughter cattle in lran[J].Front Microbiol,2014(5):536-536.
[12]胡继红,高振翔,尹铭芳.美国NCCLS2002年版抗生素药敏试验操作标准更新内容[J].中华检验医学杂志,2002(6):46-48.
[13] Sarangi L N,Priyadarshini A,Kumar S,et al.Virulence genotyping of Pasteurella multocida lsolated from multiple hosts from India[J].Sci World J,2014(2014):814109-814109.
[14]王羽,董文龙,王巍,等.牛源多杀性巴氏杆菌荚膜血清型及毒力基因检测[J].中国兽医杂志,2017,53(9):16-19.
[15]李伟杰,赵耘,杜昕波,等.产毒素多杀性巴氏杆菌的鉴定[J].中国畜牧兽医,2010,37(1):149-151.