Identification of bacterial intestinal pathogens by a PCRReverse dot blot procedure

Abstract

     Intestinal infections which are the important public health concern worldwide, are causedby the bacterial intestinal pathogens. The aim at our study is to develop a simultaneous, rapid,sensitive and specific diagnostic assay by using a combined PCR-Reverse dot blot method for theidentification of pathogen strains, including Bacillus cereus, Clostridium botulinum, Clostridiumperfringen, Staphylococcus aureus, Listeria monocytogenes, Escherichia coli O157:H7,Salmonella spp., Shigella spp., Vibrio cholerae, Vibrio parahaemolyticus, Yersinia enterocoliticaand Brucella spp.Based on the 16S and 23S DNA regions, the two sets of universal primers and twelvespecific probes were obtained for amplification and specific detection of those twelve bacterialspecies. The initial experimental results using bacterial cultures and 50 clinical samplesconfirmed the in silico hypothesis that was previously established in universal primers andprobes design as well as identified with some basic conditions for PCR and Reverse Dot Blothybridization reactions. Thus, this procedure being further tested for the other kinds of samplessuch as fecal samples or foods.