伊犁地区牛源大肠埃希菌血清分型和ERIC-PCR分型研究
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摘要
调查某肉牛养殖场和其定点屠宰加工场中牛源大肠埃希菌的表型和基因型,为产业链中大肠埃希菌的污染情况提供科学依据。对分离鉴定的牛源大肠埃希菌采用O抗原多价血清进行检测,选取其中多价4和多价5的菌株进行单价PCR(O7、O39、O5、O114、O116、O6、O9、O30、O55)的扩增,同时进行ERICPCR的基因分型。结果表明,牛源大肠埃希菌分离株以多价4、多价5、多价6和多价19较多,检出率均超过5%。单价PCR检测出O5(2株)、O114(3株)、O116(4株)、O6(6株)、O9(2株)血清型。ERIC-PCR基因分型多价4和多价5中含有13种基因型,分成6簇,同源性在59%~100%。同种多价血清在一个簇,其中D簇包括12株菌,同源性在70%以上。然而同一种血清型的不同菌株其基因型不在一个簇内。
To investigate the phenotype and genotype of Escherichia coli isolated from beef cattle farms and its designated slaughter and processing field,and to provide scientific basis for the pollution condition of E.coli in the industrial chain,the bovine E.coli was detected on the O antigen with polyvalent serum,the 4 and 5 polyvalent serum strains were selected and detected by PCR(O7,O39,O5,O114,O116,O6,O9,O30,O55)and ERIC-PCR genotyping.The results showed that 4,5,6,9 of polyvalent serotypes of the tested samples were dominant,all more than 5%.The PCR detection results were serotypes of the O5(2 strains),O114(3 strains),O116(4 strains),O6(6 strains),O9(2 strains).ERIC-PCR had 13 genotypes in 4 and 5 polyvalent serotypes,and can be divided into 6 clusters.The similarity was between 59%-100%.The same polyvalent serotype was in a cluster,the D cluster has the 12 strains and the similarity was over 70%.But the different strains in same serotype was not in same cluster genotypes.
To investigate the phenotype and genotype of Escherichia coli isolated from beef cattle farms and its designated slaughter and processing field,and to provide scientific basis for the pollution condition of E.coli in the industrial chain,the bovine E.coli was detected on the O antigen with polyvalent serum,the 4 and 5 polyvalent serum strains were selected and detected by PCR(O7,O39,O5,O114,O116,O6,O9,O30,O55)and ERIC-PCR genotyping.The results showed that 4,5,6,9 of polyvalent serotypes of the tested samples were dominant,all more than 5%.The PCR detection results were serotypes of the O5(2 strains),O114(3 strains),O116(4 strains),O6(6 strains),O9(2 strains).ERIC-PCR had 13 genotypes in 4 and 5 polyvalent serotypes,and can be divided into 6 clusters.The similarity was between 59%-100%.The same polyvalent serotype was in a cluster,the D cluster has the 12 strains and the similarity was over 70%.But the different strains in same serotype was not in same cluster genotypes.
引文
[1]郝彦龙,剡根强,王静梅,等.新疆北疆地区致犊牛腹泻大肠杆菌的分离鉴定及部分生物学特性[J].中国兽医学报,2012,32(6):848-851.
[2]庄孝飞,周秀娟,许学斌,等.2008-2012年上海市肠炎沙门氏菌分离株毒力基因筛查与ERIC-PCR分型[J].食品科学,2015,14(3):165-170.
[3]刘英玉,杨舟,吾买尔江·牙合甫,等.新疆伊犁某肉牛场大肠埃希菌及其毒力基因的检测[J].动物医学进展,2016,37(4):45-48.
[4]Atsushi I,Sunao I,Kazuko S,et al.Escherichia coli O-genotyping PCR:a comprehensive and practical platform for molecular O serogrouping[J].J Clin Microbiol,2015,53(8):2427-2432.
[5]曹芸.检测副猪嗜血杆菌的乳胶凝集试验及ERIC-PCR方法的建立与应用[D].湖北武汉:华中农业大学,2009.
[6]伍晓锋,黎毅敏,卓超,等.18株铜绿假单胞菌的耐药谱和ERIC-PCR分型[J].中国抗生素杂志,2007,32(9):560-563.
[7]韩海红.生食贝类中副溶血性弧菌污染水平调查、定量风险评估和分离菌株特征分析[D].北京:中国疾病预防控制中心,2015.
[8]王文豪,苏丹萍,张丹琳,等.华南地区致病性猪大肠杆菌的鉴定和血清型分布[J].中国兽医杂志,2013,49(4):33-34.
[9]李永丽,应春妹,陈艺升.ERIC-PCR技术在鲍曼不动杆菌基因分型中的应用评估[J].检验医学,2013,28(7):621-624.
[10]Fendri I,Hassena A B,Grosset N,et al.Genetic diversity of food-isolated Salmonellastrains through pulsed field gel electrophoresis(PFGE)and enterobacterial repetitive intergenic consensus(ERIC-PCR)[J].PLoS One,2013,8(12):e81315-e81315.
[11]王秋艳.三种基因分型技术在病原微生物溯源方面的应用[D].广东广州:华南理工大学,2010.
[12]Warriner K,Aldsworth T G,Kaur S,et al.Cross-contamination of carcasses and equipment during pork processing[J].J Appl Microbiol,2002,93(1):169-177.
[13]Wang H,Shu R,Zhao Y,et al.Analysis of ERIC-PCR genomic polymorphism of Salmonellaisolates from chicken slaughter line[J].Eur Food Res Technol,2014,239(3):543-548.
[14]高宇,贾丹,田甜,等.湖北地区91株嗜水气单胞菌ERIC-PCR指纹图谱分型[J].云南农业大学学报:自然科学版,2016,31(6):1022-1030.
[2]庄孝飞,周秀娟,许学斌,等.2008-2012年上海市肠炎沙门氏菌分离株毒力基因筛查与ERIC-PCR分型[J].食品科学,2015,14(3):165-170.
[3]刘英玉,杨舟,吾买尔江·牙合甫,等.新疆伊犁某肉牛场大肠埃希菌及其毒力基因的检测[J].动物医学进展,2016,37(4):45-48.
[4]Atsushi I,Sunao I,Kazuko S,et al.Escherichia coli O-genotyping PCR:a comprehensive and practical platform for molecular O serogrouping[J].J Clin Microbiol,2015,53(8):2427-2432.
[5]曹芸.检测副猪嗜血杆菌的乳胶凝集试验及ERIC-PCR方法的建立与应用[D].湖北武汉:华中农业大学,2009.
[6]伍晓锋,黎毅敏,卓超,等.18株铜绿假单胞菌的耐药谱和ERIC-PCR分型[J].中国抗生素杂志,2007,32(9):560-563.
[7]韩海红.生食贝类中副溶血性弧菌污染水平调查、定量风险评估和分离菌株特征分析[D].北京:中国疾病预防控制中心,2015.
[8]王文豪,苏丹萍,张丹琳,等.华南地区致病性猪大肠杆菌的鉴定和血清型分布[J].中国兽医杂志,2013,49(4):33-34.
[9]李永丽,应春妹,陈艺升.ERIC-PCR技术在鲍曼不动杆菌基因分型中的应用评估[J].检验医学,2013,28(7):621-624.
[10]Fendri I,Hassena A B,Grosset N,et al.Genetic diversity of food-isolated Salmonellastrains through pulsed field gel electrophoresis(PFGE)and enterobacterial repetitive intergenic consensus(ERIC-PCR)[J].PLoS One,2013,8(12):e81315-e81315.
[11]王秋艳.三种基因分型技术在病原微生物溯源方面的应用[D].广东广州:华南理工大学,2010.
[12]Warriner K,Aldsworth T G,Kaur S,et al.Cross-contamination of carcasses and equipment during pork processing[J].J Appl Microbiol,2002,93(1):169-177.
[13]Wang H,Shu R,Zhao Y,et al.Analysis of ERIC-PCR genomic polymorphism of Salmonellaisolates from chicken slaughter line[J].Eur Food Res Technol,2014,239(3):543-548.
[14]高宇,贾丹,田甜,等.湖北地区91株嗜水气单胞菌ERIC-PCR指纹图谱分型[J].云南农业大学学报:自然科学版,2016,31(6):1022-1030.