Evaluation of Polymerase Chain Reaction for Detecting Coliform Bacteria in Drinking Water Sources

Isfahani, Bahram Nasr; Fazeli, Hossein; Babaie, Zeinab; Poursina, Farkhondeh; Moghim, Sharareh; Rouzbahani, Meisam

Evaluation of Polymerase Chain Reaction for Detecting Coliform Bacteria in Drinking Water Sources

Document Type : Original Article

Authors

Department of Microbiology and Virology, Isfahan University of Medical Sciences, Isfahan, Iran

Abstract

Background: Coliform bacteria are used as indicator organisms for detecting fecal pollution in water. Traditional methods including microbial culture tests in lactose-containing media and enzyme-based tests for the detection of β-galactosidase; however, these methods are time-consuming and less specific. The aim of this study was to evaluate polymerase chain reaction (PCR) for detecting coliform. Materials and Methods: Totally, 100 of water samples from Isfahan drinking water source were collected. Coliform bacteria and Escherichia coli were detected in drinking water using LacZ and LamB genes in PCR method performed in comparison with biochemical tests for all samples. Results: Using phenotyping, 80 coliform isolates were found. The results of the biochemical tests illustrated 78.7% coliform bacteria and 21.2% E. coli. PCR results for LacZ and LamB genes were 67.5% and 17.5%, respectively. Conclusion: The PCR method was shown to be an effective, sensitive, and rapid method for detecting coliform and E. coli in drinking water from the Isfahan drinking water sources.

Keywords

References

1.	Fatemeh D, Reza ZM, Mohammad A, Salomeh K, Reza AG, Hossein S, et al. Rapid detection of coliforms in drinking water of Arak city using multiplex PCR method in comparison with the standard method of culture (Most Probably Number). Asian Pac J Trop Biomed 2014;4:404-9.
2.	Stepenuck KF, Wolfson LG, Liukkonen BW, Iles JM, Grant TS. Volunteer monitoring of E. coli in streams of the upper Midwestern United States: A comparison of methods. Environ Monit Assess 2011;174:625-33.
3.	Fatahi-Bafghi M. Phenotypic and molecular identification of nocardia in brain abscess. Adv Biomed Res 2017;6:49. [PUBMED] [Full text]
4.	Awwa A. Standard Methods for the Examination of Water and Wastewater. Washington, DC: American Public Health Association; 1998. p. 20.
5.	Rompré A, Servais P, Baudart J, de-Roubin MR, Laurent P. Detection and enumeration of coliforms in drinking water: Current methods and emerging approaches. J Microbiol Methods 2002;49:31-54.
6.	McFeters GA, Cameron SC, LeChevallier MW. Influence of diluents, media, and membrane filters on detection fo injured waterborne coliform bacteria. Appl Environ Microbiol 1982;43:97-103.
7.	Tharannum S, Sunitha S, Nithya J, Chandini M, Vanitha J, Manjula T, et al. Molecular confirmation of the presence of coliforms in drinking water using polymerase chain reaction. Kathmandu Univ J Sci Eng Technol 2009;5:130-6.
8.	Kämpfer P, Nienhüser A, Packroff G, Wernicke F, Mehling A, Nixdorf K, et al. Molecular identification of coliform bacteria isolated from drinking water reservoirs with traditional methods and the Colilert-18 system. Int J Hyg Environ Health 2008;211:374-84.
9.	Clifford RJ, Milillo M, Prestwood J, Quintero R, Zurawski DV, Kwak YI, et al. Detection of bacterial 16S rRNA and identification of four clinically important bacteria by real-time PCR. PLoS One 2012;7:e48558.
10.	Bej AK, Steffan RJ, DiCesare J, Haff L, Atlas RM. Detection of coliform bacteria in water by polymerase chain reaction and gene probes. Appl Environ Microbiol 1990;56:307-14.
11.	Ahmed OB, Asghar AH, Elhassan MM. Comparison of three DNA extraction methods for polymerase chain reaction (PCR) analysis of bacterial genomic DNA. Afr J Microbiol Res 2014;8:598-602.
12.	Spellman FR, Drinan JE. The Drinking Water Handbook. McNeese State University: CRC Press; 2012.
13.	Tantawiwat S, Tansuphasiri U, Wongwit W, Wongchotigul V, Kitayaporn D. Development of multiplex PCR for the detection of total coliform bacteria for Escherichia coli and Clostridium perfringens in drinking water. Southeast Asian J Trop Med Public Health 2005;36:162-9.
14.	Van Belkum A. DNA fingerprinting of medically important microorganisms by use of PCR. Clin Microbiol Rev 1994;7:174-84.
15.	Locas A, Barthe C, Margolin AB, Payment P. Groundwater microbiological quality in Canadian drinking water municipal wells. Can J Microbiol 2008;54:472-8.
16.	Al-Thwani AN. Monitoring of drinking water quality in Baghdad city by using polymerase chain reaction (PCR). J Babylon Univ Pure Appl Sci 2014;3.
17.	Clinical and Laboratory Standards Institute. Performance Standards for Antimicrobial Susceptibility Testing, Twenty-Second Informational Supplement. CLSI Document M100-S22. Wayne, Pennsylvania, USA: Clinical and Laboratory Standards Institute; 2012.
18.	Abo-Amer AE, Soltan el-SM, Abu-Gharbia MA. Molecular approach and bacterial quality of drinking water of urban and rural communities in Egypt. Acta Microbiol Immunol Hung 2008;55:311-26.
19.	Spierings G, Ockhuijsen C, Hofstra H, Tommassen J. Polymerase chain reaction for the specific detection of Escherichia coli/Shigella. Res Microbiol 1993;144:557-64.
20.	Tsen HY, Lin CK, Chi WR. Development and use of 16S rRNA gene targeted PCR primers for the identification of Escherichia coli cells in water. J Appl Microbiol 1998;85:554-60.

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