OPTIMIZATION OF MOLECULAR DETECTION FOR Vibrio cholerae and PATHOGENIC Escherichia coli USING MULTIPLEX PCR

Authors

  • Diana Elizabeth Waturangi Atma Jaya Catholic University
  • Jason Petrus Atma Jaya Catholic University
  • Rico Kosasih Atma Jaya Catholic University
  • Felicia Roseline Atma Jaya Catholic University

DOI:

https://doi.org/10.36547/be.299

Keywords:

Rapid detection, PCR multiplex, Foodborne diseases, Vibrio cholerae, Pathogenic Escherichia coli

Abstract

Vibrio cholerae and pathogenic Escherichia coli 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 V. cholerae and pathogenic E. coli 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 V. cholerae and pathogenic E. coli were obtained with a primer concentration of 16 µM for ctxA and ompU, 30 µM for ace, and 50 µM for zot, and toxR; 2 µM for elt and 5 µM for stx, 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 V. cholerae and pathogenic E. coli respectively. The optimized method was qualified to be used as a molecular detection for V. cholerae as well as EHEC and ETEC detection according to ISO/TS 20836 (2017)  from drinking water samples.

Downloads

Published

2021-05-24

How to Cite

Waturangi, D. E., Petrus, J., Kosasih, R., & Roseline, F. (2021). OPTIMIZATION OF MOLECULAR DETECTION FOR Vibrio cholerae and PATHOGENIC Escherichia coli USING MULTIPLEX PCR. Bacterial Empire, e299. https://doi.org/10.36547/be.299

Issue

Section

Bacteriology Articles
Received 2021-04-14
Accepted 2021-05-18
Published 2021-05-24