Real-Time PCR assay and melting curve analysis for identification of diarrheagenic <em>Escherichia coli</em> in paediatric population

Abstract

In developing countries, including India, diarrhea causes considerable death among infants and children. Early diagnosis plays a vital role in preventing the fatality rate. Conventional methods for early detection of diarrheagenic <em>E. coli</em> (DEC) are complicated and inaccurate. Hence, in this study we proposed a syber green based Real-Time PCR to determine the prevalence of DEC. Detection of diarrheagenic <em>E. coli </em> using Real-Time Syber Green based PCR assay was performed in 120 isolates including 80 isolates from healthy children. Virulent genes were detected in 106 (88.3%) samples [39 (97.5%) and 67 (83.7%)] in cases and controls, respectively. EPEC was found (<em>typical</em>, <em> atypical </em> and <em>bfp</em>) in 87 (72.5%) specimens followed by EAEC (<em>eagg</em>) in 76 (63.3%), ETEC (<em>elt </em> and e<em>st</em>) in 55 (45.8%) and EHEC (<em>stx</em> and <em>hyla</em>) in 11 (9.2%) specimens. Melting curve analysis showed distinct peaks for all target genes.