Multiplex PCR assay for direct detection of diarrheagenic escherichia coli from clinical stool specimens

ABSTRACT

Background: To distinguish the different types of pathogenic E. coli with other non-pathogenic E.coli in the intestine is extremely important in diagnosis. Up to date there are at least six types of E. coli that causes diarrhea. Objectives: We have designed a multiplex PCR assay for the direct detection of 6 categories of diarrheagenic Escherichia coli. Subjects and method: This techniques proved to be specific and rapid for detecting virulence genes from Shiga toxin-producing (stx and eae), enteropathoogenic (eae), enterotoxigenic (elt, est), ennteroinvasive (ipaH), enteroaggregative (aggR), and diffuse adherent (daaE) Esscherichia coli. The technique was applied to 295 clinical stool specimens. Results: The highest prevalence is EAggEC with 51 positive samples.(17.29%), 48 EIEC (16.27%), 17 EPEC (5.76%), 8 ETEC (LT) (2.71%), 5 ETEC (ST) (1.69%), 1 DAEC (0.34%), no STEC positive and 19 mix infections (6.44%). Conclusion: Multiplex PCR assay is a quick and highly accuurate technique. It is not only specific but can also amplify 7 virulence genes of diarrrheagenic E.coli at the same time. This method would offer an effective alternative to traditional culture methods for the identification and differentiation of human diarrhaegenic Escherichia coli.