Establishment of Multiplex DPO-PCR Method for Detection of Shiga Toxin-producing Escherichia coli O26
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Abstract:
In order to develop a triple DPO-PCR assay for the detection of Shiga toxin-producing Escherichia coli O26 in foods based DPO primers, three sets of DPO primers were designed according to shiga toxin stx1, stx2 and O antigen wzx O26 genes. Their specificity, sensitivity and detection effect were evaluated. The results showed that an optimized system was established, which showed a high specificity and sensitivity. The amplification could product at 49~69 ℃. The detection limit of this assay for Escherichia coli O26 was 3.8×103 cfu/g. Further, this method demonstrated good detection results in artifical contaminated samples and natural food samples. The triple DPO-PCR assay in this work based on DPO primers possessed highly advantages of high efficiency, specificity and wide annealing temperature range, which can provide a reliable and efficient molecule assay for the rapid and accurate detection of Shiga toxin-producing E. coli O26 in food samples method.
