Escherichia coli O157 is an important food-borne pathogen. 39 E. coli O157 strains identified during 2014 and 2015 in retail foods in china were identified by double PCR to understand the biological characteristics of food-borne E. coli O157. The virulence genes and antimicrobial susceptibility of isolates were also detected, and the genetic diversity of Escherichia coli O157 was analyzed by Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR) typing. The results showed that 8 were identified as O157:H7 and 31 were O157 among the 39 strains. , Eae gene was detected in all the isolates, and stx2 was in 82.50% strains of isolates, while stx1 was not detected in the tested 11 virulence genes. Other virulence genes such as espA, etpD, tir, toxB, iha, and katP were detected with the carrying rate of 92.31%, 94.87%, 87.18%, 79.49%, 69.23%, and 46.15%, respectively. Antimicrobial susceptibility results indicated that the isolates strains were highly resistant to TE, SXT, S, C, and AMP. More than 30% of the isolates were resistance to three and more antibiotics. The CRISPR typing indicated that these isolates strains had a high genetic diversity. Thirty-three of the 39 strains had CRISPR1 loci, including 8 E. coli O157:H7 and 26 E. coli O157. The 8 E. coli O157:H7 produced the same spacer maps, while 26 E. coli O157 produced 13 diversified spacer spectra. This study can provide important basic data for food borne disease surveillance, disease traceability and epidemiological research.
[1] Galli L, Brusa V, Singh P, et al. High prevalence of clade 8 Escherichia coli O157:H7 isolated from retail meat and butcher shop environment [J]. Infection Genetics and Evolution, 2016, 45: 1-5
[2] D'Astek B A, DEL-Castillo L L, Miliwebsky E, et al. Subtyping of Escherichia coli O157:H7 strains isolated from human infections and healthy cattle in Argentina [J]. Foodborne Pathogens and Disease, 2012, 9(5): 457-464
[3] Sharapov U M, Wendel A M, Davis J P, et al. Multistate outbreak of Escherichia coli O157:H7 infections associated with consumption of fresh spinach: United States, 2006 [J]. Journal of Food Protection, 2016, 79(12): 2024-2030
[4] 陈雅君,王亚宾,张莉娟,等.动物源性肠出血性大肠杆菌O157:H7及其3个毒力基因的多重PCR快速检测研究[J].中国人兽共患病学报,2013,29(7):686-691
CHEN Ya-jun, WANG Ya-bin, ZHANG Li-juan, et al. Detection of animal-derived Escherichia coli O157:H7 and its three virulence genes by multiplex PCR technique [J]. Chinese Journal of Zoonoses, 2013, 29(7): 686-691
[5] 张书萧.大肠杆菌O157的分子流行病学调查和毒力因子研究[D].长春:吉林农业大学,2012
ZHANG Shu-xiao. Molecular epidemiology and study on virulence factor of Escherichia coli O157 [D]. Changchun: Jilin Agricultural University, 2012
[6] 万成松.大肠杆菌O157:H7的毒力岛与毒力因子的研究进展[J].微生物学免疫学进展,2006,34(2):58-62
WAN Cheng-song. The pathogenic islands and virulence factors of Escherichia coli O157:H7 [J]. Progress in Microbiology and Immunology, 2006, 34(2): 58-62
[7] Kim H S, Chon J W, Kim Y J, et al. Prevalence and characterization of extended-spectrum-β-lactamase- producing Escherichia coli, and Klebsiella pneumoniae, in ready-to-eat vegetables [J]. International Journal of Food Microbiology, 2015, 207: 83-86
[8] Yang B, Wang Q, Cui S, et al. Characterization of extended-spectrum beta-lactamases-producing Salmonella strains isolated from retail foods in Shaanxi and Henan province, China [J]. Food Microbiology, 2014, 42(12): 14-18
[9] Ahmed A M, Shimamoto T. Molecular analysis of multidrug resistance in Shiga toxin-producing Escherichia coli O157:H7 isolated from meat and dairy products [J]. International Journal of Food Microbiology, 2015, 193: 68-73
[10] 叶菊莲,占利,梅玲玲,等.利用MLST技术对浙江省大肠杆菌O157的分子流行病学研究[J].中国人兽共患病学报,2011,27(10):901-904
YE Ju-lian, ZHAN Li, MEI Ling-ling, et al. Multilocus sequence typing analysis application in molecular epidemiologicalresearch of Escherichia coli O157 isolates in Zhejiang province [J]. Chinese Journal of Zoonoses, 2011, 27(10): 901-904
[11] 王开,裴志花,陈中秋,等.常见食源性致病菌及其分子分型技术的研究进展[J].中国兽医杂志,2013,49(12):51-54
WANG Kai, PEI Zhi-hua, CHEN Zhong-qiu, et al. Research progress of common food-borne pathogens and their molecular typing techniques [J]. Chinese Journal of Veterinary Medicine, 2013, 49(12): 51-54
[12] Fabre L, Zhang J, Guigon G, et al. CRISPR typing and subtyping for improved laboratory surveillance of Salmonella infections [J]. PLoS One. 2012, 7(5): e36995
[13] Shariat N, Dudley E G. CRISPRs: molecular signatures used for pathogen subtyping [J]. Applied and Environmental Microbiology, 2014, 80(2): 430-439
[14] Di H, Ye L, He Y, et al. Comparative analysis of CRISPR loci in different Listeria monocytogenes, lineages [J]. Biochemical and Biophysical Research Communications, 2014, 454(3): 399-403
[15] 梁文娟,张荣光,段广才,等.基于CRISPR对大肠埃希菌O157:H7的检测[J].西安交通大学学报(医学版),2016, 37(5):748-753
LIANG Wen-juan, ZHANG Rong-guang, DUAN Guang-cai, et al. Detection of Escherichia coli O157:H7 based on CRISPR locu [J]. Department of Preventive Medicine, Henan University of Science and Technology, 2016, 37(5): 748-753
[16] Monaghan á, Byrne B, Fanning S, et al. Serotypes and virulence profiles of atypical enteropathogenic Escherichia coli, (EPEC) isolated from bovine farms and abattoirs [J]. Journal of Applied Microbiology, 2013, 114(2): 595-603
[17] Paton A W, Paton J C. Detection and characterization of Shiga toxigenic Escherichia coli by using multiplex PCR assays for stx1, stx2, eaeA, enterohemorrhagic E. coli hlyA, rfbO111, and rfbO157 [J]. Journal of Clinical Microbiology, 1998, 36(2): 598-602
[18] 水新云,王虎虎,高峰,等.4株E. coli O157:H7毒力基因检测及其冷应激损伤[J].食品科学,2016,37(4):176-180
SHUI Xin-yun, WANG Hu-hu, GAO Feng, et al. Detection of virulence genes of four Escherichia coli O157:H7 strains and cold stress injury in the bacteria [J]. Food Science, 2016, 37(4): 176-180
[19] 白莉,刘秀梅,付萍,等.2005~2007年中国食品中疑似O157大肠埃希菌的鉴定及毒素基因的检测[J].卫生研究,2010,39(3):335-338
BAI Li, LIU Xiu-mei, FU Ping, et al. Serotyping and virulence genes of suspected Escherichia coli O157 strains in food from 2005 to 2007 [J]. Journal of Hygiene Research, 2010, 39(3): 335-338
[20] Sallam K I, Mohammed M A, Ahdy A M, et al. Prevalence, genetic characterization and virulence genes of sorbitol-fermenting Escherichia coli O157:H- and E. coli O157:H7 isolated from retail beef [J]. International Journal of Food Microbiology, 2013, 165(3): 295-301
[21] Dong P, Zhu L, Mao Y, et al. Prevalence and characterization of Escherichia coli O157:H7 from samples along the production line in Chinese beef-processing plants [J]. Food Control, 2015, 54(1): 39-46
[22] 赵嘉咏,穆玉姣,张白帆,等.河南省2009-2010年出血性大肠埃希菌O157毒力基因分布与脉冲场凝胶电泳分型研究[J].中华流行病学杂志,2015,36(11):1324-1326
ZHAO Jia-yong, MU Yu-jiao, ZHANG Bai-fan, et al. Distribution of virulence genes and PFGE molecular typing of Entero-hemorrhagic Escherichia coli O157 in Henan from 2009 to 2010 [J]. Chinese Journal of Epidemiology, 2015, 36(11): 1324-1326
[23] Msolo L, Igbinosa E O, Okoh A I. Prevalence and antibiogram profiles of Escherichia coli O157:H7 isolates recovered from three selected dairy farms in the Eastern Cape Province, South Africa [J]. Asian Pacific Journal of Tropical Disease, 2016, 6(11): 990-995
[24] Disassa N, Sibhat B, Mengistu S, et al. Prevalence and antimicrobial susceptibility pattern of E.coli O157:H7 isolated from traditionally marketed raw cow milk in and around Asosa town, Western Ethiopia [J]. Veterinary Medicine International, 2017, 2017: 7581531
[25] Beyi A F, Fite A T, Tora E, et al. Prevalence and antimicrobial susceptibility of Escherichia coli O157 in beef at butcher shops and restaurants in central Ethiopia [J]. Bmc Microbiology, 2017, 17(1): 49
[26] Ison S A, Delannoy S, Bugarel M, et al. Genetic diversity and pathogenic potential of attaching and effacing Escherichia coli O26:H11 strains recovered from bovine feces in the United States [J]. Applied and Environmental Microbiology, 2015, 81(11): 3671-8
[27] Tymensen L D. CRISPR1 analysis of naturalized surface water and fecal Escherichia coli suggests common origin [J]. Microbiology Open, 2016, 5(3): 527-533
[28] 庄孝飞.基于CRISPR序列的沙门氏菌分子分型方法的建立[D].上海:上海交通大学,2015
ZHUANG Xiao-fei. Development of CRISPR-Based molecular typing method of Salmonella [D]. Shanghai: Shanghai Jiao Tong University, 2015