Multiplex polymerase chain reaction for identification of six diarrheagenic Escherichia coli and Shigella

Objective To develop a rapid and specific multiplex polymerase chain reaction (PCR) system for the identification of six diarrheagenic Escherichia coli and Shigella. Methods The multiplex PCR system was established based on 9 virulence genes eae, stx1, stx2, elt, estIa, estIb, aggR, ipaH, afaD and 16S rRNA gene rrs was used as control. The multiplex PCR system was optimized and evaluated by using 174 diarrheagenic E. coli strains, 24 Shigella strains and 44 other enteric pathogen strains. Results The multiplex PCR could amplify the related virulence genes, and the results were 100% consistent with those of the simplex PCR. The sensitivity of the multiplex PCR was 4.56 101 CFU/reaction or 1.14 104 CFU/ml for pure culture. Conclusion The established multiplex PCR system is specific for screening and detection of six diarrheagenic E. coli pathovars and Shigella.