TY - JOUR AU - Waturangi, Diana Elizabeth AU - Petrus, Jason AU - Kosasih, Rico AU - Roseline, Felicia PY - 2021/08/13 Y2 - 2024/03/29 TI - OPTIMIZATION OF MOLECULAR DETECTION FOR Vibrio cholerae and PATHOGENIC Escherichia coli USING MULTIPLEX PCR JF - Bacterial Empire JA - BE VL - 4 IS - 3 SE - Bacteriology Articles DO - 10.36547/be.299 UR - https://office.scicell.org/index.php/BE/article/view/299 SP - e299 AB - <p><em>Vibrio cholerae</em> and pathogenic <em>Escherichia coli</em> were considered as main causative agent foodborne diseases especially in many developing countries, such as Indonesia. Thereby, rapid detection of these pathogenic bacteria is necessary to treat food-borne related diseases causing by these bacteria. In this case, multiplex PCR allows multiple genes amplification in one reaction thereby enable to perform rapid detection of these pathogenic bacteria. The objective of this study is to optimize uniplex and multiples PCR of <em>V. cholerae</em> and pathogenic <em>E. coli</em> detection and determine the sensitivity and specificity of this assays. We used various virulence genes for each pathogenic bacterium as markers for uniplex and multiplex PCR detection. Based on this research, the optimum results of <em>V. cholerae</em> and pathogenic <em>E. coli</em> were obtained with a primer concentration of 16 µM for <em>ctxA</em> and <em>ompU</em>, 30 µM for <em>ace</em>, and 50 µM for <em>zot</em>, and <em>toxR</em>; 2 µM for <em>elt</em> and 5 µM for <em>stx</em>, respectively. Finally, based on the standardization method by ISO/TS 20836 these assays had 0% false positive, 0% false negative, 100% specificity, and 100% sensitivity; 0% false positive, 4% false negative, 100% specificity, and 96% sensitivity for <em>V. cholerae</em> and pathogenic <em>E. coli</em> respectively. The optimized method was qualified to be used as a molecular detection <em>for V. cholerae</em> as well as EHEC and ETEC detection according to ISO/TS 20836 (2017)  from drinking water samples.</p> ER -