Genotyping analysis on foodborn Listeria monocytogenes starins isolated in Beijing region
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摘要:
目的 对北京地区肉类产品中单增李斯特菌进行全基因组测序,了解不同序列型分布特点及不同菌株间的亲缘关系。 方法 采用高通量测序技术对110株单增李斯特菌进行全基因组测序、多位点序列分型(MLST)分析,采用相关软件对测序数据进行基因组装、基因预测与功能注释。采用抗性基因数据库(CARD)、毒力因子数据库(VFDB)筛选耐药基因、致病基因。 结果 110株分离菌株全基因组测序MLST分型共分为17个ST型,其中1株菌为ST新型(STnew1),ST9为优势型。新型STnew1具有大环内酯类、氨基糖苷类、恶唑烷酮类、氯霉素类、四环素类、甲氧苄氨嘧啶类等多种耐药基因,缺失基因prfA、hpt、plcA、plcB、inlC、hlyA。其中20.9 % 的菌株均携带耐药基因。除6株无害李斯特菌外,其余菌株均携带单增李斯特菌毒力岛(LIPI-1)。 结论 肉类食品中单增李斯特菌存在多种耐药基因,毒力基因分布以毒力岛(LIPI-1)为主,具有潜在的致病性;相关部门应加强肉类及肉类制品的监管,进一步降低单增李斯特菌所致食源性疾病流行风险。 Abstract:Objective To examine genotype distribution of Listeria monocytogenes (Lm) strains isolated from meat product specimens in Beijing and genetic correlations among the strains with whole genome sequencing (WGS). Methods Totally 110 Lm strains were sequenced with WGS and analyzed with multilocus sequence typing (MLST). Gene assembly, prediction and function annotation for WGS results of the strains were performed using relevant softwares. The Comprehensive Antibiotic Research Database (CARD) and the Virulence Factors Database (VFDB) were used to screen drug resistance genes and pathogenic genes of the isolated Lm strains. Results The 110 Lm strains were classified into 17 sequence type (ST) types; among the 17 ST types, there was a unknown ST type (namely STnew1) and ST9 was the dominant ST type. The STnew1 has multiple drug resistance genes including genes against macrolides, aminoglycosides, oxazolidinones, chloramphenicols, tetracyclines, and trimethoprim and gene deletion of prfA, hpt, plcA, plcB, inlC, and hlyA. Of the 110 Lm strains, all carried the Listeria monocytogenes pathogenicity island (LIPI-1) gene, except for 6 non-pathogenic strains, and 20.9% carried drug resistance genes. Conclusion Multiple drug resistance genes are prevalence in Listeria monocytogenes strains isolated from meat products and LIPI-1 gene is dominant among the drug resistance genes. The results suggest that contamination of Listeria monocytogenes in meat products needs to be monitored and controlled to reduce the risk of foodborne diseases caused by the bacteria. -
Key words:
- whole genome sequencing /
- Listeria monocytogenes /
- genotyping /
- virulence gene /
- drug resistance gene
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表 1 109株单增李斯特菌MLST型分布
ST型别 菌株数 丰度(%) ST9 40 36.7 ST121 16 14.7 ST155 12 11.0 ST8 9 8.3 ST87 8 7.3 ST378 4 3.7 ST120 4 3.7 ST542 3 2.8 ST537 2 1.8 ST59 2 1.8 ST602 1 0.9 ST101 1 0.9 ST196 2 1.8 ST11 1 0.9 ST705 2 1.8 ST2 1 0.9 ST307 1 0.9 表 2 单增李斯特菌耐药基因状况
样品(菌株号) 耐药基因 ST分型 鸡翅(L21) aac(6′) -Iaa、aph(3″) -Ib aph(6) -Id、blaTEM– 1B、fosX、sul2、tet(M) ST155 猪肘(L25) fosX、Tet(M)、dfrG ST155 三黄鸡(L33) aac(6′) -Iaa aph(3″) -Ib aph(6) -Id、blaTEM– 1B、fosX fosA7、qnrS1、sul3、tet(A)、dfrA14 ST121 鸡胸(L35) aac(6′) -Iaa、aph(3′) -III、Str、fosX、lsa(A)、tet(M) ST307 鸭腿(L52) aph(3′) -III、str、fosX、erm(B)、lsa(A)、optrA、cat、fexB、tet(L)、tet(M)tet(O/W/32/O)、dfrG ST121 散养鸡(L53) aph(3′) -III、str、erm(B)、lsa(A)、optrA、cat、fexB、tet(L)、tet(M)、tet(O/W/32/O)、dfrG STnew1 鸭胸(L63) FosX、lsa(A)、tet(L)、VanC4XY ST121 鸡胸(L128) aadA3、blaCTX– M– 148、blaCTX– M– 65、blaSHV– 12、blaSHV– 13、blaSHV– 129、blaSHV– 155、blaSHV– 172 blaSHV– 31、fosA6、fosA、fosX、qnrS1 ST378 表 3 李斯特菌毒力基因缺失状况
样品(菌株号) 毒力基因缺失情况 MLST分型 猪里脊(L80) Mpl、actA、prfA、plcB、plcA、bsh、hpt、fpbA、inlC ST602 鸡大胸(L99) Mpl、actA、prfA、plcB、plcA、bsh、hpt ST542 牛肉(L89) Mpl、actA、prfA、plcB、plcA、bsh、hpt ST537 猪五花肉(L46) Mpl、actA、prfA、plcB、plcA、bsh、hpt ST542 三黄鸡(L104) Mpl、actA、prfA、plcB、plcA、bsh、hpt ST542 牛肉(L124) Mpl、actA、prfA、plcB、plcA、bsh、hpt ST537 散养鸡(L53) prfA、hpt、plcA、plcB、inlC、hlyA STnew1 -
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