Abstract:
Bacterial pathogens continue to be the most common cause of enteric infections that leads to diarrhoea especially in poor-resource settings and is rated by the World Health Organization as the second contributory factor in the mortality and morbidity of children. Diagnostic techniques for detection of these pathogens is mainly through culture that takes long and may not often useful be for instituting immediate treatment. Multiplex polymerase chain reaction (mPCR) is a technique which enables simultaneous amplification of many targets of interest in one reaction by using more than one pair of primers. This study was aimed at isolating and identifying enteropathogens in children under five years of age attending Mbagathi District Hospital, Nairobi, Kenya One hundred stool samples from the children under the age of five years with diarrhoea were collected in polypots and used for the study. The stool samples were enriched on Selenite F and then subcultured on XLD and MacConkey agar. Biochemical tests were carried out to further identify the bacterial species. For evaluation of the mPCR technique, the Seeplex Diarrhoea-B1 ACE detection was used in a multiplex assay that permitted simultaneous amplification of target DNA of Salmonella spp. (S. bongori and S. enterica), Shigella spp. (S. flexneri, S. boydii, S. sonnei and S. dysenteriae), Vibrio spp. (V. cholerae, V. parahamolyticus and V. vulnificus), Clostridium difficile toxin B, Campylobacter spp. (C. jejuni and C. coli) and Internal Control (IC). Out of the 100 samples that were analysed in this study Salmonella spp, Shigella spp and E. coli (VTEC) showed the highest prevalence of 73%. This was followed by Vibrio spp at 26%, then Campylobacter spp at 6%. The study found mPCR to be relatively easy to perform, reproducible and rapid, therefore recommends it in rapid disease diagnosis and timely decision making for treatment of infection.
