东莞市区肉菜市场熟肉制品中金黄色葡萄球菌污染溯源分析
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摘要
目的优化熟肉制品中金黄色葡萄球菌的快速污染溯源分析方法。方法对熟肉制品及其加工环境进行采样,应用3MPerifilmTM金黄色葡萄球菌快速测试片对样品定量检测,对分离出的阳性菌株运用全自动微生物基因指纹鉴定系统RiboPrinter,结合EcoRI酶,对其进行分子溯源。结果 168个样品中分离出的7株菌共分为2种核糖体基因型,其中6株为同一核糖体条带类型,显示来源于相同克隆株。结论该方法全程只需3 d时间,快速稳定,可满足监管快速检测鉴定、分型溯源的要求。RiboPrinter的优势在于快速溯源,进行种的鉴定时应结合其他手段来进行确认。
Objective To optimize the rapid method for source analysis of Staphylococcus aureus in cooked meat product. Methods Staphylococcus aureus were collected from the meat products and the processing environment by using the 3 M PerifilmTM. The isolations were source analyzed by the automated ribotyping system using the restriction enzymes EcoRI. Results The 7 strains isolated from 168 samples were divided into 2 ribosome genotypes, and 6 of which were of the same ribosome band type, indicating that they were derived from the same clone. Conclusion This method only needs 3 days for the whole process, which is fast and stable, and can meet the requirements of supervision, rapid detection, identificationand typing traceability. The advantage of RiboPrinter is the rapid traceability. The identification of the species should be combined with other means.
Objective To optimize the rapid method for source analysis of Staphylococcus aureus in cooked meat product. Methods Staphylococcus aureus were collected from the meat products and the processing environment by using the 3 M PerifilmTM. The isolations were source analyzed by the automated ribotyping system using the restriction enzymes EcoRI. Results The 7 strains isolated from 168 samples were divided into 2 ribosome genotypes, and 6 of which were of the same ribosome band type, indicating that they were derived from the same clone. Conclusion This method only needs 3 days for the whole process, which is fast and stable, and can meet the requirements of supervision, rapid detection, identificationand typing traceability. The advantage of RiboPrinter is the rapid traceability. The identification of the species should be combined with other means.
引文
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[3]Croucher NJ,Finkelstein JA,Pelton SI,et al.Population genomics of post-vaccine changes in pneumococcal epidemiology[J].Nat Genet,2013,45(6):656-663.
[4]Didelot X,Pang B,Zhou ZM,et al.The role of China in the global spread of the current Cholera Pandemic[J].PLoS Genet,2015,11(3):e1005072.
[5]Holt KE,Wertheim H,Zadoks RN,et al.Genomic analysis of diversity,population structure,virulence,and antimicrobial resistance in Klebsiella pneumoniae,an urgent threat to public health[J].Proc Natl Acad Sci USA,2015,112(27):3574-3581.
[6]微生物实验室国家食品安全风险评估中心.国家食源性致病微生物全基因组测序数据库及测序分析平台建设进展[EB/OL].[2018-02-11].http://www.cfsa.net.cn/Article/News.aspx?id=9E132A0E2B5EC262635B068CC82F649F328B1F9F1CEBB65F.China National Center For Food Safety Risk Assessment.Progress of the national foodborne pathogenic microorganisms whole genome sequencing database and sequencing analysis platform[EB/OL].[2018-02-11].http://www.cfsa.net.cn/Article/News.aspx?id=9E132A0E2B5EC262635B068CC82F649F328B1F9F1CEBB65F.
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[8]Inglis TJ,O’Reilly L,Foster N,et al.Comparison of automated ribotyping and DNA macrorestriction analysis of Burkholderia pseudomaller[J].JClin Micro-Bil,2002,40(9):3198-3203.
[9]苏妙贞,曾晓琮,韩志杰,等.金黄色葡萄球菌定量检测中四种培养基的比较[J].食品安全导刊,2017,(7):139-140.Su MZ,Zeng XC,Han ZJ,et al.Comparison of four cultures for quantitative detection of staphylococcus aureus[J].Chin Food Saf Magaz,2017,(7):139-140.
[10]周少君,梁辉,朱海明,等.广东省熟肉制品中金黄色葡萄球菌的污染调查及初步风险分析[J].中国食品卫生杂志,2014,26(5):496-450.Zhou SJ,Liang H,Zhu HM,et al.Staphylococcus aureus contamination and risk evaluation in ready-to-eat meat products in Guangdong province[J].Chin J Food Hyg,2014,26(5):496-450.
[11]许振伟,韩奕奕,孟瑾,等.熟食肉制品中金黄色葡萄球菌风险评估基础研究[J].包装与食品机械,2012,30(5):40-43.Xu ZW,Han YY,Meng J,et al.Basic research on risk assessment of Staphylococcus aureus in meat products[J].Packag Food Mach,2012,30(5):40-43.
[12]容冬丽,吴清平,吴诗,等.我国部分地区即食食品和蔬菜中金黄色葡萄球菌污染分布及耐药和基因分型情况[J].微生物学报,2018,58(2):314-323.Rong DL,Wu QP,Wu S,et al.Prevalence,antimicrobial susceptibility,and genetic characteristics of Staphylococcus aureus from retail ready-to-eat foods and vegetables insome regions of China[J].Acta Microbiol Sin,2018,58(2):314-323.
[13]刘思超,罗泽燕,徐励琴,等.惠州市熟肉制品中金黄色葡萄球菌污染状况及PFGE分型[J].中国热带医学,2018,18(5):459-463.Liu SC,Luo ZY,Xu LQ,et al.Contamination of Staphylococcus aureusin cooked meat products and molecular typing by PFGE in Huizhou[J].Chin Trop Med,2018,18(5):669-671.
[14]朱海明,邓峰,严纪文,等.r DNA图谱在金黄色葡萄球菌引起食物中毒溯源中的应用[J].中国热带医学,2007,7(5):669-671.Zhu HM,Deng F,Yan JW,et al.Application of rDNA fringerprinting of Staphylococcus aureus in tracing the source of microbial food-poison-thing[J].Chin Trop Med,2007,7(5):669-671.
[15]刘培,杨思,良娟,等.金黄色葡萄球菌分离株自动化核糖体分型的研究[J].食品研究与开发,2016,37(1):138-141.Liu P,Yang S,Liang J,et al.Automated Ribotyping of Staphylococcus aureus Monocytogenes[J].Food Res Dev,2016,37(1):138-141.
[16]董健,郑晶,林杰,等.3M PetrifilmTM金黄色葡萄球菌检验测试片的评价[J].食品与发酵科技,2014,50(4):56-60.Dong J,Zheng J,Lin J,et al.Validation of 3M petrifilmTM Staphylococcus aureus count plate[J].Food Ferment Technol,2014,50(4):56-60.
[17]钟玮,冯震,鲍英,等.金黄色葡萄球菌的API-TAPH生化鉴定及核糖体分型分析[J].中国抗生素杂志,2013,38(5):354-359.Zhong W,Feng Z,Bao Y,et al.Identification of Staphylococcus aureus by API-STAPH test and ribosomal typing analysis[J].Chin J Antibiot,2013,38(5):354-359.
[18]杨永刚,陈瑜,陈松,等.耐甲氧西林金黄色葡萄球菌基因分型测定技术进展[J].中华医院感染学杂,2016,6(26):1433-1436.Yang YG,Chen Y,Chen S,et al.Current status fogenotyping technologies of methicillin-resistant Staphylococcus aureus[J].Chin J Nosocomiol,2016,6(26):1433-1436.
[19]GB 4789.10-2016食品安全国家标准食品微生物学检验金黄色葡萄球菌检验[S].GB 4789.10-2016 National food safety standard-Detection of Staphylococcus aureus in food[S].
[2]闻剑,梁辉,戴光伟,等.2008-2011年广东省熟肉制品中常见食源性致病菌污染状况分析[J].中国卫生食品杂志,2013,25(1):68-70.Wen J,Liang H,Dai GW,et al.Analysis of food-borne pathogen contaminationin ready-to-eat meat products in Guangdong province from2008 to 2011[J].Chin J Food Hyg,2013,25(1):68-70.
[3]Croucher NJ,Finkelstein JA,Pelton SI,et al.Population genomics of post-vaccine changes in pneumococcal epidemiology[J].Nat Genet,2013,45(6):656-663.
[4]Didelot X,Pang B,Zhou ZM,et al.The role of China in the global spread of the current Cholera Pandemic[J].PLoS Genet,2015,11(3):e1005072.
[5]Holt KE,Wertheim H,Zadoks RN,et al.Genomic analysis of diversity,population structure,virulence,and antimicrobial resistance in Klebsiella pneumoniae,an urgent threat to public health[J].Proc Natl Acad Sci USA,2015,112(27):3574-3581.
[6]微生物实验室国家食品安全风险评估中心.国家食源性致病微生物全基因组测序数据库及测序分析平台建设进展[EB/OL].[2018-02-11].http://www.cfsa.net.cn/Article/News.aspx?id=9E132A0E2B5EC262635B068CC82F649F328B1F9F1CEBB65F.China National Center For Food Safety Risk Assessment.Progress of the national foodborne pathogenic microorganisms whole genome sequencing database and sequencing analysis platform[EB/OL].[2018-02-11].http://www.cfsa.net.cn/Article/News.aspx?id=9E132A0E2B5EC262635B068CC82F649F328B1F9F1CEBB65F.
[7]周海健,阚飙.细菌基因组分型方法的应用研究进展[J].疾病监测,2016,31(8):668-674.Zhou HJ,Kan B.Progress in application research of bacterium genome-based subtyping[J].Dis Surv,2016,31(8):668-674.
[8]Inglis TJ,O’Reilly L,Foster N,et al.Comparison of automated ribotyping and DNA macrorestriction analysis of Burkholderia pseudomaller[J].JClin Micro-Bil,2002,40(9):3198-3203.
[9]苏妙贞,曾晓琮,韩志杰,等.金黄色葡萄球菌定量检测中四种培养基的比较[J].食品安全导刊,2017,(7):139-140.Su MZ,Zeng XC,Han ZJ,et al.Comparison of four cultures for quantitative detection of staphylococcus aureus[J].Chin Food Saf Magaz,2017,(7):139-140.
[10]周少君,梁辉,朱海明,等.广东省熟肉制品中金黄色葡萄球菌的污染调查及初步风险分析[J].中国食品卫生杂志,2014,26(5):496-450.Zhou SJ,Liang H,Zhu HM,et al.Staphylococcus aureus contamination and risk evaluation in ready-to-eat meat products in Guangdong province[J].Chin J Food Hyg,2014,26(5):496-450.
[11]许振伟,韩奕奕,孟瑾,等.熟食肉制品中金黄色葡萄球菌风险评估基础研究[J].包装与食品机械,2012,30(5):40-43.Xu ZW,Han YY,Meng J,et al.Basic research on risk assessment of Staphylococcus aureus in meat products[J].Packag Food Mach,2012,30(5):40-43.
[12]容冬丽,吴清平,吴诗,等.我国部分地区即食食品和蔬菜中金黄色葡萄球菌污染分布及耐药和基因分型情况[J].微生物学报,2018,58(2):314-323.Rong DL,Wu QP,Wu S,et al.Prevalence,antimicrobial susceptibility,and genetic characteristics of Staphylococcus aureus from retail ready-to-eat foods and vegetables insome regions of China[J].Acta Microbiol Sin,2018,58(2):314-323.
[13]刘思超,罗泽燕,徐励琴,等.惠州市熟肉制品中金黄色葡萄球菌污染状况及PFGE分型[J].中国热带医学,2018,18(5):459-463.Liu SC,Luo ZY,Xu LQ,et al.Contamination of Staphylococcus aureusin cooked meat products and molecular typing by PFGE in Huizhou[J].Chin Trop Med,2018,18(5):669-671.
[14]朱海明,邓峰,严纪文,等.r DNA图谱在金黄色葡萄球菌引起食物中毒溯源中的应用[J].中国热带医学,2007,7(5):669-671.Zhu HM,Deng F,Yan JW,et al.Application of rDNA fringerprinting of Staphylococcus aureus in tracing the source of microbial food-poison-thing[J].Chin Trop Med,2007,7(5):669-671.
[15]刘培,杨思,良娟,等.金黄色葡萄球菌分离株自动化核糖体分型的研究[J].食品研究与开发,2016,37(1):138-141.Liu P,Yang S,Liang J,et al.Automated Ribotyping of Staphylococcus aureus Monocytogenes[J].Food Res Dev,2016,37(1):138-141.
[16]董健,郑晶,林杰,等.3M PetrifilmTM金黄色葡萄球菌检验测试片的评价[J].食品与发酵科技,2014,50(4):56-60.Dong J,Zheng J,Lin J,et al.Validation of 3M petrifilmTM Staphylococcus aureus count plate[J].Food Ferment Technol,2014,50(4):56-60.
[17]钟玮,冯震,鲍英,等.金黄色葡萄球菌的API-TAPH生化鉴定及核糖体分型分析[J].中国抗生素杂志,2013,38(5):354-359.Zhong W,Feng Z,Bao Y,et al.Identification of Staphylococcus aureus by API-STAPH test and ribosomal typing analysis[J].Chin J Antibiot,2013,38(5):354-359.
[18]杨永刚,陈瑜,陈松,等.耐甲氧西林金黄色葡萄球菌基因分型测定技术进展[J].中华医院感染学杂,2016,6(26):1433-1436.Yang YG,Chen Y,Chen S,et al.Current status fogenotyping technologies of methicillin-resistant Staphylococcus aureus[J].Chin J Nosocomiol,2016,6(26):1433-1436.
[19]GB 4789.10-2016食品安全国家标准食品微生物学检验金黄色葡萄球菌检验[S].GB 4789.10-2016 National food safety standard-Detection of Staphylococcus aureus in food[S].