Optimization of multiplex PCR conditions for rapid detection of Escherichia coli O157:H7 virulence genes
ثبت نشده
چکیده
The main source of E. coli 0157:H7 is cattle, but recent studies showed high percentage of outbreaks contributed by contaminated water. The occurrence of E. coli O157:H7 in environmental water samples poses a potential threat to human health. The aim of this study was to establish a protocol for the detection of the pathogen E. coli O157:H7 and E. coli virulence genes (eaeA, rfbE, hly, stx1, and stx2) in a multiplex PCR protocol using six specific primer pairs. The target genes produced species-specific amplicons at 625 bp, 397 bp, 296 bp, 166 bp, 210 bp and 484 bp for E. coli O157:H7 (fliCh7 gene) and virulence genes (eaeA, rfbE, hly, stx1, and stx2) respectively. The results obtained show that the established PCR protocol is suitable for a rapid and specific analysis of the pathogenic E. coli O157:H7 in environmental water samples for the assessment of microbiological risks.
منابع مشابه
Isolation, Characterization and Antibiotic Resistance of Shiga Toxin-Producing Escherichia coli in Hamburger and Evolution of Virulence Genes stx1, stx2, eaeA and hly by Multiplex PCR
Background & Objectives: Shiga toxin-producing Escherichia coli (STEC) O157:H7 have emerged as pathogens that can cause food-borne infections and severe and potentially fatal illnesses in humans. E.coli O157:H7 colonizes the digestive tract of cattle and is transmitted to humans by food and water. The objectives of this study were to characterize the prevalence of E.coli O157:H7 isolates in ham...
متن کاملPrevalence of Virulence Genes of Escherichia Coli O157:H7 Isolated from Patients with Urinary Tract Infections in Shiraz, Iran
Abstract Background and Objective: Escherichia coli O157:H7 is one of the most well-known pathogenic bacteria worldwide that can develop severe diseases such as hemolytic uremic syndrome (HUS). This study aimed to assess the prevalence of virulence genes of E. coli O157:H7 in patients with suspected urinary tract infections (UTIs). Material and Methods: This cross-sectional study was co...
متن کاملSemi-automated fluorogenic PCR assays (TaqMan) forrapid detection of Escherichia coli O157:H7 and other shiga toxigenic E. coli.
Semi-automated detection of Enterohaemorrhagic Escherichia coli (EHEC) O157:H7 and non-O157:H7 Shiga toxin-producing E. coli (STEC) was achieved using fluorogenic polymerase chain reaction (PCR). These PCR assays were designed to amplify 80, 120 and 150 bp regions of virulence genes stx1, stx2 and eaeA, respectively, using specific primers. The fluorogenic probes were used for specific detectio...
متن کاملDetection of Shiga toxin-producing Escherichia coli (STEC) in faeces of healthy calves in Mashhad, Iran
The aim of this study was to identify virulent Shiga toxin-producing Escherichia coli (STEC) strains isolated from faecal samples of 100 clinically healthy calves. In the present study, a total of 100 Escherichia coli (E. coli) isolates from clinically healthy calves belonging to 6 different farms located in Khorasan Razavi province, Iran, were examined for presence of virulence genes character...
متن کاملIdentification of shiga toxin producing Escherichia coli O157:H7 in raw cow milk samples from dairy farms in Mashhad using multiplex PCR assay
In this study 130 bulk tank milk samples which were delivered to the Pegah Pasturisation Factory inMashhad were collected randomly during the summer months. Samples were firstly enriched in modifiedtrypticase soy broth containing novobiocin, followed by plating onto sorbitol MacConkey agar supplementedwith cefixime and potassium tellurite for isolation of Escherichia coli O157:H7. Consequently ...
متن کامل