Abstract

COMPUTATION AND IN-SILICO VALIDATION OF CANDIDATE PRIMERS FOR PCR-FREE ON-SITE DETECTION OF ENTEROTOXIGENIC ESCHERICHIA COLI

Rajesh Singh Tomar and Anurag Jyoti†

Abstract

Over population, industrialization and urban crowding has led to the pollution of water resources resulting into the outbreaks of diarrheal diseases, threatening human lives. Enterotoxigenic Escherichia coli (ETEC) is a potential pathovar responsible for water-borne diarrhea among children and travelers. Rapid and on-site detection of ETEC is important for the management of these diseases. This creates a selection pressure on the scientists to develop a potential detection tool to diagnose the causative agent. The present study attempts to design and in-silico validate the specific candidate primers of isothermal amplification to detect ETEC. The primers for isothermal amplification were computed targeting LT1 signature gene of ETEC. A web-based dedicated software Primer Explorer V4 was used to compute the primers. All the primers were further checked for the inclusivity and exclusivity in nucleotide-BLAST. Results showed that all the primers are highly specific towards their respective targets. Therefore, the present study implicates the computation of primers of PCR-free isothermal amplification for the detection of ETEC.

Keywords: Enterotoxigenic Escherichia coli, Nanoarray, Isothermal amplification, point-of-care diagnostics.