广州市58株单增李斯特菌菌株特征分析
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摘要
目的了解广州市动物源性食品中单增李斯特菌菌株优势血清型、携带毒力基因和分子分型情况。方法对广州地区365份动物源性食品中的单增李斯特菌分离鉴定,并进行血清分型,毒力基因鉴定和肠道细菌基因间重复序列扩增(ERIC-PCR)。结果单增李斯特菌阳性率为15.89%(58/365),其中生畜肉类检出率最高,为25.38%(33/130);血清型分析将58株单增李斯特菌分为4种血清型,分别为1/2b、1/2a、1/2c和4b,优势的血清型为1/2b(44.80%)和1/2a(32.80%);10株缺失毒力基因actA;ERIC-PCR扩增出9～18条100～8 000 bp之间的条带,将58株单增李斯特菌在相似系数为0.8处分为6个群19个类型。结论广州市动物源性食品中单增李斯特菌菌株污染严重,优势血清型是与人或动物感染有关的致病血清型(1/2b和1/2a),大部分毒力基因均存在,ERIC-PCR结果显示分子型别呈多样性,应加强防控。
Objective To analyze the phenotypic and gene characteristics of the Listeria monocytogenes isolated from animal-derived food samples in Guangzhou.Methods Presumptive Listeria monocytogenes were isolated from 365 animal-derived food samples using the national food safety standard,Food Microbiological Examination: Listeria monocytogenes(GB/T 4789.30-2016).The serological classification,virulence genes,and Enterobacterial repetitive intergenic consensus sequence-PCR(ERIC-PCR) were performed to provide valuable strain discrimination.Results 58 strains of Listeria monocytogenes were detected in the 365 animal-derived food samples.The total detection rate was 15.89%(58/365),with the highest detection rate(25.38%,33/130) in raw livestock meat.The serotype 1/2 b(44.83%,26/58) and 1/2 a(32.76%,19/58) were dominant,followed by serotype 1/2 c(18.97%,11/58) and 4 b(3.44%,2/58). And all were positive for inlA,inlC,inlJ,plcA,prfA,actA,hlyA and Iap gene except ten stains that lacked the actA gene.ERIC-PCR revealed that the strains isolated from animal-derived food samples had distinct PCR fingerprints.Conclusion The study demonstrates the prevalence of pathogenic Listeria monocytogenes in the animal-derived food samples.The predominated serotype 1/2 b and 1/2 a and most virulence-related genes were detected in the Listeria monocytogenes strains,which seems to have serious public health consequences.
Objective To analyze the phenotypic and gene characteristics of the Listeria monocytogenes isolated from animal-derived food samples in Guangzhou.Methods Presumptive Listeria monocytogenes were isolated from 365 animal-derived food samples using the national food safety standard,Food Microbiological Examination: Listeria monocytogenes(GB/T 4789.30-2016).The serological classification,virulence genes,and Enterobacterial repetitive intergenic consensus sequence-PCR(ERIC-PCR) were performed to provide valuable strain discrimination.Results 58 strains of Listeria monocytogenes were detected in the 365 animal-derived food samples.The total detection rate was 15.89%(58/365),with the highest detection rate(25.38%,33/130) in raw livestock meat.The serotype 1/2 b(44.83%,26/58) and 1/2 a(32.76%,19/58) were dominant,followed by serotype 1/2 c(18.97%,11/58) and 4 b(3.44%,2/58). And all were positive for inlA,inlC,inlJ,plcA,prfA,actA,hlyA and Iap gene except ten stains that lacked the actA gene.ERIC-PCR revealed that the strains isolated from animal-derived food samples had distinct PCR fingerprints.Conclusion The study demonstrates the prevalence of pathogenic Listeria monocytogenes in the animal-derived food samples.The predominated serotype 1/2 b and 1/2 a and most virulence-related genes were detected in the Listeria monocytogenes strains,which seems to have serious public health consequences.
引文
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[3] 中华人民共和国卫生部.GB/T 4789.30-2010 食品安全国家标准食品卫生微生物学检验单核细胞增生李斯特氏菌检验[S].北京:中国标准出版社,2010.
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[5] Liu D,Lawrence ML,Austin FW,et al.A multiplex PCR for species-and virulence-specific determination of Listeria monocytogenes[J].Journal of Microbiology Methods,2007,71(2):133-140.
[6] 马爱静,王艳,王毅,等.北京市一些地区生肉标本中单增李斯特菌的分离及其分子流行病学特征分析[J].中国人兽共患病学报,2015,31(5):403-407.
[7] 李秀娟,赵冬,潘琢,等.29种毒力基因在91株食源性单核细胞增生李斯特氏菌中的分布[J].中国人兽共患病学报,2017,33(11):972-978.
[8] 宋筱瑜,裴晓燕,徐海滨,等.我国零售食品单增李斯特菌污染的健康风险分级研究[J].中国食品卫生杂志,2015,27(4):447-450.
[9] Wieczorek K,Dmowska K,Osek J.Prevalence,characterization,and antimicrobial resistance of listeria monocytogenes isolates from bovine hides and carcasses[J].Applied Environmental Microbiology,2012,78(6):2043-2045.
[10] Vázquez-Boland JA,Domínguez-Bernal G,González-Zorn B,et al.Pathogenicity islands and virulence evolution in Listeria[J].Microbes and Infection,2001,3(7):571-584.
[2] Cole MB,Jones MV,Holyoak C.The effect of pH,salt concentration and temperature on the survival and growth of Listeria monocytogenes[J].The Journal of Applied Bacteriology,1990,69:63-72.
[3] 中华人民共和国卫生部.GB/T 4789.30-2010 食品安全国家标准食品卫生微生物学检验单核细胞增生李斯特氏菌检验[S].北京:中国标准出版社,2010.
[4] Soni DK,Singh M,Singh DV,et al.Virulence and genotypic characterization of Listeria monocytogenes isolated from vegetable and soil samples[J].BMC Microbiology,2014,14:241.
[5] Liu D,Lawrence ML,Austin FW,et al.A multiplex PCR for species-and virulence-specific determination of Listeria monocytogenes[J].Journal of Microbiology Methods,2007,71(2):133-140.
[6] 马爱静,王艳,王毅,等.北京市一些地区生肉标本中单增李斯特菌的分离及其分子流行病学特征分析[J].中国人兽共患病学报,2015,31(5):403-407.
[7] 李秀娟,赵冬,潘琢,等.29种毒力基因在91株食源性单核细胞增生李斯特氏菌中的分布[J].中国人兽共患病学报,2017,33(11):972-978.
[8] 宋筱瑜,裴晓燕,徐海滨,等.我国零售食品单增李斯特菌污染的健康风险分级研究[J].中国食品卫生杂志,2015,27(4):447-450.
[9] Wieczorek K,Dmowska K,Osek J.Prevalence,characterization,and antimicrobial resistance of listeria monocytogenes isolates from bovine hides and carcasses[J].Applied Environmental Microbiology,2012,78(6):2043-2045.
[10] Vázquez-Boland JA,Domínguez-Bernal G,González-Zorn B,et al.Pathogenicity islands and virulence evolution in Listeria[J].Microbes and Infection,2001,3(7):571-584.