Molecular screening and characterization of Shiga toxin-producing Escherichia coli in retail foods

Abstract

Shiga toxin-producing Escherichia coli (STEC) strains are one of the most important recently emerged groups of food borne pathogens. This study investigated the prevalence of molecular markers for STEC and characteristics of E. coli O157 isolates from foods sold at retail markets in Wuhan, China. A total of 489 samples (350 meat products and 139 raw vegetables) were purchased from 22 large scale markets between July of 2011 and September of 2013. The meat samples consisted of frozen chicken products, raw pork, raw beef, frozen fish products and processed duck products. The raw vegetable samples consisted of lettuce, bok choy, radish, spinach, cucumber, and tomato. Shiga toxin genes (stx1 and stx2) and an O-group marker of the seven main pathogenic STEC serogroups (O157, O26, O45, O103, O111, O121, and O145) were detected in the samples by using PCR. 100% agreement was obtained between the results of the PCR targeting for wzyO157 and the PCR targeting for rfbEO157 gene. The result demonstrated that PCR assay targeting for wzyO157 gene can be employed as an effective screening method for E. coli O157 in food sample. In the study, E. coli O157 and non-O157 STEC were detected in 55 (11.2%) and 75 (15.3%) samples by PCR screening, respectively. There was significant difference in the occurrence of STEC contamination between supermarkets (19/127, 15.0%) and open markets (111/362, 30.7%) (P < 0.05). Out of 489 samples, 5 samples carried O45, 1 sample carried O145 and 1 sample carried O111. Markers for O103, O26 and O121 were not detected. This result differed from other reports. Immunomagnetic separation based cultivation technique was used to isolate E. coli O157 from 27 food samples collected in 2013. Finally 7 E. coli O157 isolates were obtained. Among the 7 isolates, the prevalent stx genotype was stx1a and stx2a. Four E. coli O157 strains exhibited toxic effects on Vero cells, while 3 isolates had no detectable cytotoxicity effects even though they contained stx genes. All E. coli O157 isolates were sensitive to the 12 antimicrobials tested except for roxithromycin. There are some inconsistencies between the PCR screening and culture results. Characteristics of STEC isolates should be evaluated and considered for monitoring STEC contamination in foods.