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

Document Type : Original Article


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


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.


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.  Back to cited text no. 1
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.  Back to cited text no. 2
Fatahi-Bafghi M. Phenotypic and molecular identification of nocardia in brain abscess. Adv Biomed Res 2017;6:49.  Back to cited text no. 3
[PUBMED]  [Full text]  
Awwa A. Standard Methods for the Examination of Water and Wastewater. Washington, DC: American Public Health Association; 1998. p. 20.  Back to cited text no. 4
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.  Back to cited text no. 5
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.  Back to cited text no. 6
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.  Back to cited text no. 7
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.  Back to cited text no. 8
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.  Back to cited text no. 9
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.  Back to cited text no. 10
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.  Back to cited text no. 11
Spellman FR, Drinan JE. The Drinking Water Handbook. McNeese State University: CRC Press; 2012.  Back to cited text no. 12
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.  Back to cited text no. 13
Van Belkum A. DNA fingerprinting of medically important microorganisms by use of PCR. Clin Microbiol Rev 1994;7:174-84.  Back to cited text no. 14
Locas A, Barthe C, Margolin AB, Payment P. Groundwater microbiological quality in Canadian drinking water municipal wells. Can J Microbiol 2008;54:472-8.  Back to cited text no. 15
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.  Back to cited text no. 16
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.  Back to cited text no. 17
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.  Back to cited text no. 18
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.  Back to cited text no. 19
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.  Back to cited text no. 20