Culture-Free Detection of Enterotoxigenic Escherichia Coli in Food by Polymerase Chain Reaction
Journal Title: International Journal for Research in Applied Science and Engineering Technology (IJRASET) - Year 2015, Vol 3, Issue 4
Abstract
Microbial quality of food has immense importance for protection of human health in rural and urban settings. Rapid and specific detection of enterotoxigenic Escherichia coli (ETEC) is critical for the management of the food-borne diarrheal diseases threatening human lives. Conventional methods for detection of ETEC involve enrichment and biochemical identification which are laborious, time consuming and less sensitive. Polymerase Chain Reaction, using specific primers is a powerful molecular technique for rapid and specific detection of ETEC. The present study aims to detect ETEC bearing virulent signature gene LT1 in food samples. Street fruit juices like Citrus (Citrus limetta), Pineapple (Ananas comosus), Sugarcane (Saccharum spp.) were procured from local market. Apart from these, vegetables used for garnishing the dishes such as Fenugreek (Trigonella foenum-graecum) Mint (Mentha spp.) and Coriander (Coriandrum sativum) leaves were purchased from local market. Multigenomic DNA templates were prepared by boiling prep and purified using Sodium acetate-ethanol method. Genomic DNA from pure cultures was also prepared and purified. This was followed by PCR targeting the LT1 gene to detect ETEC in food samples. Food samples collected from local market were analysed and found contaminated with ETEC. The amplified products from samples’ DNA were same as compared to positive control. The amplification based assay developed here is rapid with high specificity for detection of ETEC in food samples.
Authors and Affiliations
Anurag Jyoti, Mohit Agarwal, Rajesh Singh Tomar
Keywords
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