Detection of Enterohemorrhagic Escherichia coli (EHEC) in Consumption Water Source using Multiplex PCR Method

Khoirun Nihayati, Yuanita Rachmawati, Saiku Rokhim, Linda Prasetyaning Widayanti

2018

Abstract

Escherichia coli is microorganism that often causes diarrhea. One of the most pathogenic E. coli bacteria for humans is Escherichia coli O157: H7. These bacteria are included in E. coli Enterohemorrhagic (EHEC) bacteria which produce shiga like toxin. Shiga like toxin causes Hemorrhagic Colitis (HC) and Hemolytic Ureamic Syndrome (HUS). Water is a basic need for living things. Water that is suitable for use as a source of consumption must fullfill certain chemical, physical and biological requirements. Biologically, the source of consumption water should not contain E. coli. The presence of these bacteria indicates fecal contamination. Water, especially consumption water, is a potential source for the spread of microorganisms that cause infection, including E. coli O157: H7 bacteria. The purpose of this research is to detect Escherichia coli O157: H7 bacteria in consumption water source sample using multiplex PCR method. The primers used in this study were fliCh7 (625 bp), rfbE (296 bp), stx1 (210 bp), stx2 (484 bp), eaeA (397 bp), and hly (166 bp) genes. The process of bacterial isolation was carried out through several stages, namely presumptive test, confirmed test, and completed test. Isolation of bacterial DNA was carried out using boiling cell method. Amplification process was carried out in conditions of 94˚C-2 minutes, followed by 35 cycles of 94˚C-20 seconds, 63˚C-1 minute annealing, and 72˚C-1 extension minutes, and post-extension 72˚C-10 minutes. Next, 2% agarose gel electerophoresis was done at 50 volts for 65 minutes. The results showed that from the total 7 samples tested, all are positive samples of coliform bacteria, 6 are positive samples of Escherichia coli bacteria, and 3 are positive samples of Escherichia coli O157: H7 bacteria with virulent expression of different genes including stx1 and eaeA genes. An analysis of all irrigation systems is needed to determine the entry of coliform contamination, E. coli and E. coli O157: H7

Paper Citation

in Harvard Style

Nihayati K., Rachmawati Y., Rokhim S. and Widayanti L. (2018). Detection of Enterohemorrhagic Escherichia coli (EHEC) in Consumption Water Source using Multiplex PCR Method. In Proceedings of the Built Environment, Science and Technology International Conference - Volume 1: BEST ICON, ISBN 978-989-758-414-5, pages 108-112. DOI: 10.5220/0008907000002481

in Bibtex Style

@conference{best icon18,
author={Khoirun Nihayati and Yuanita Rachmawati and Saiku Rokhim and Linda Prasetyaning Widayanti},
title={Detection of Enterohemorrhagic Escherichia coli (EHEC) in Consumption Water Source using Multiplex PCR Method},
booktitle={Proceedings of the Built Environment, Science and Technology International Conference - Volume 1: BEST ICON,},
year={2018},
pages={108-112},
publisher={SciTePress},
organization={INSTICC},
doi={10.5220/0008907000002481},
isbn={978-989-758-414-5},
}

in EndNote Style

TY - CONF

JO - Proceedings of the Built Environment, Science and Technology International Conference - Volume 1: BEST ICON,
TI - Detection of Enterohemorrhagic Escherichia coli (EHEC) in Consumption Water Source using Multiplex PCR Method
SN - 978-989-758-414-5
AU - Nihayati K.
AU - Rachmawati Y.
AU - Rokhim S.
AU - Widayanti L.
PY - 2018
SP - 108
EP - 112
DO - 10.5220/0008907000002481