Bai Y, Wang W, Yan L, Yang S R, Yan S F, Dong Y P, Zhao B C, Zhao Y Y, Xu J, Hu Y J, Li F Q
Key Lab of Food Safety Risk Assessment, Ministry of Health, China National Center for Food Safety Risk Assessment, Beijing 100021, China.
Zhonghua Yu Fang Yi Xue Za Zhi. 2018 Apr 6;52(4):364-371. doi: 10.3760/cma.j.issn.0253-9624.2018.04.007.
To analyses the antimicrobial resistance and molecular characterization of 21 MRSA isolates cultured from retail foods from different provinces in China, and evaluate the molecular typing methods. Twenty-one MRSA isolates were obtained from national foodborne pathogen surveillance network in 2012 (Chinese salad, 3; milk, 1; cake, 2; rice, 1; cold noodle, 1; spiced beef, 1; dumpling, 1; packed meal, 1; salad, 1; raw pork, 9). The antimicrobial resistance of 21 strains to 12 antimicrobial agents was tested by broth dilution method. Polymerase chain reaction (PCR) and DNA sequencing were performed to obtain the genetic types of MLST (ST) and spa typing. The clonal complex (CC) was assigned by eBURST soft and the MLVA type (MT) and MLVA complex (MC) were identified via the database of the MLVA website (http://www.mlva.net). I pulsed-field gel electrophoresis (Ⅰ-PFGE) was also carried out to obtain the PFGE patterns of 21 strains. The genetic diversity and discriminatory power of typing were calculated by the Simpson's index of diversity (diversity index, DI) to find out the best genotyping method for MRSA. All MRSA isolates showed multi-drug resistance(MDR), and were resistant to oxacillin, benzylpenicillin, clindamycin and erythromycin, and 71.4% (15/21), 47.6% (10/21), 42.9% (9/21) and 28.6% (6/21) of the MRSA isolates were resistant to tetracycline, ciprofloxacin, trimethoprim/sulfamethoxazole and gentamicin, respectively. Moreover, one strain was found to be resistant to all three antimicrobials of levofloxacin, moxifloxacin and rifampicin. Great diversity was found in these food-associated MRSA (6 STs, 7 types, and 9 MTs). PFGE patterns were more diverse than those of other three molecular typing methods (19 pulse types). The index of diversity (DI) of PFGE, MLVA, typing and MLST was 0.99, 0.80, 0.73, and 0.61, respectively. Among the MRSA isolates, CC9-ST9-t899-MT929-MC2236 (PFGE Cluster Ⅴ) was the most prevalent clone, which were all cultured from raw pork (9 isolates). Besides, two MRSA were identified as CC59-ST338-t437-MT621-MC621 (PFGE Cluster Ⅳ). Different clone had their own resistance spectrum profiles. The food-borne MRSA isolates were all MDR in this study. Different clones had their own resistance spectrum profiles. MLVA represented a promising tool for molecular epidemiology tracing of MRSA in foodborne disease events.
分析从中国不同省份零售食品中培养的21株耐甲氧西林金黄色葡萄球菌(MRSA)的耐药性和分子特征,并评估分子分型方法。2012年从国家食源性病原体监测网络获得21株MRSA(中式沙拉3株、牛奶1株、蛋糕2株、米饭1株、冷面1株、五香牛肉1株、饺子1株、盒饭1株、沙拉1株、生猪肉9株)。采用肉汤稀释法检测21株菌株对12种抗菌药物的耐药性。进行聚合酶链反应(PCR)和DNA测序以获得多位点序列分型(MLST,ST)的基因类型和spa分型。通过eBURST软件确定克隆复合体(CC),并通过MLVA网站(http://www.mlva.net)的数据库鉴定MLVA类型(MT)和MLVA复合体(MC)。还进行了脉冲场凝胶电泳(Ⅰ-PFGE)以获得21株菌株的PFGE图谱。通过辛普森多样性指数(多样性指数,DI)计算分型的遗传多样性和鉴别力,以找出MRSA的最佳基因分型方法。所有MRSA分离株均表现出多重耐药(MDR),对苯唑西林、苄青霉素、克林霉素和红霉素耐药,分别有71.4%(15/21)、47.6%(10/21)、42.9%(9/21)和28.6%(6/21)的MRSA分离株对四环素、环丙沙星、甲氧苄啶/磺胺甲恶唑和庆大霉素耐药。此外,发现1株对左氧氟沙星、莫西沙星和利福平这三种抗菌药物均耐药。这些与食品相关的MRSA存在很大的多样性(6种ST型、7种类型和9种MT型)。PFGE图谱比其他三种分子分型方法的图谱更多样化(19种脉冲类型)。PFGE、MLVA、分型和MLST的多样性指数(DI)分别为0.99、0.80、0.73和0.61。在MRSA分离株中,CC9-ST9-t899-MT929-MC2236(PFGE聚类Ⅴ)是最常见的克隆型,均从生猪肉中培养得到(9株分离株)。此外,鉴定出2株MRSA为CC59-ST338-t437-MT621-MC621(PFGE聚类Ⅳ)。不同克隆具有各自的耐药谱特征。本研究中食源性MRSA分离株均为MDR。不同克隆具有各自的耐药谱特征。MLVA是食源性疾病事件中MRSA分子流行病学溯源的一种有前景的工具。