Application Of The Multiplex Polymerase Chain Reaction (M-PCR) For The Screening Of Vibrio Spp. From Rivers In Kuching, Sarawak

Abstract

The present study was conducted to investigate the occurrence of Vibrio spp. from selected rivers in Kuching,Sarawak (Malaysia) using Multiplex Polymerase Chain Reaction (m-PCR). During the 6-month study period,19 samples were collected monthly from 7 rivers, followed by simultaneous detection of three Vibrio spp.,Vibrio parahaemolyticus, Vibrio cholerae and Vibrio vulnificus, in a single tube PCR reaction. Three sets ofprimers targeting the thermolabile (tl), outer membrane protein (ompW) and hemolysin/cytolysin genes(vulCulsl) of V. parahaemolyticus, V. cholerae and V. vulnificus, respectively, were used. The results indicatedthat V. parahaemolyticus was the predominant species, occurring approximately 60.9% throughout thesampling period, followed by V. cholerae (23.1%) and V. vulnificus (16.0%). The months of July andDecember were found to be the months where all three Vibrio spp. were found to be at higher frequencies inthe river samples. Results analyzed also indicated that the rivers with the highest prevalence of the three Vibriospp. were Tambak Sejingkat, followed by Sungai Jernang and Sungai Tabuan. We conclude that m-PCR is apowerful and useful tool for the rapid and simultaneous detection of V. parahaemolyticus, V. cholerae and V.vulnificus from the riverine environments without the need for isolation and culturing. Furthermore, thismethod is highly specific, and could be applied in diagnostic laboratories for larger scale epidemiologicalinvestigations of Vibrio spp.