Volume 21, Issue 5 (Iranian South Medical Journal 2018)
Iran South Med J 2018, 21(5): 383-392 |
Back to browse issues page
1- Department of Microbiology and Parasitology, School of Medicine, Bushehr University of Medical Sciences, Bushehr, Iran
2- Department of Microbiology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran
3- The Persian Gulf Marine Biotechnology Research Center, The Persian Gulf Biomedical Sciences Research Institute, Bushehr University of Medical Sciences, Bushehr, Iran , mahaghighy@gmail.com
2- Department of Microbiology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran
3- The Persian Gulf Marine Biotechnology Research Center, The Persian Gulf Biomedical Sciences Research Institute, Bushehr University of Medical Sciences, Bushehr, Iran , mahaghighy@gmail.com
Abstract: (3335 Views)
Background: Standard methods of identification have not been able to solve all issues concerning E. coli. With the development of genomic studies, PCR appears promising to deal with the shortcomings. This study aimed to utilize PCR with specific primers for lacZ, uidA, cyd, and lacY gene segments to identify
environmental E. coli isolates.
Materials and methods: PCR and the aforementioned four primers were used for molecular identification of E. coli on purified genome DNA from 120 environmental E. coli isolates, standard strains of Shigella, and Enterohaemorrhagic E. coli strain as controls. All environmental E. coli isolates were isolated from Bushehr coastal areas and identified in a previous study by standard bacteriological methods and then
preserved in -70 ˚C for further studies.
Results: The primers successfully showed their ability to identify the targets in environmental isolates and standard strains. It is shown that the four PCR fragments related to lacZ, uidA, cyd, and lacY genes were observed only for E. coli isolates and strains.
Conclusion: PCR method proved capable to distinguish E. coli from Shigella as the most phylogenetically related genus and contrary to the classical methods, it could detect enterohaemorrhagic strains as Escherichia coli.
environmental E. coli isolates.
Materials and methods: PCR and the aforementioned four primers were used for molecular identification of E. coli on purified genome DNA from 120 environmental E. coli isolates, standard strains of Shigella, and Enterohaemorrhagic E. coli strain as controls. All environmental E. coli isolates were isolated from Bushehr coastal areas and identified in a previous study by standard bacteriological methods and then
preserved in -70 ˚C for further studies.
Results: The primers successfully showed their ability to identify the targets in environmental isolates and standard strains. It is shown that the four PCR fragments related to lacZ, uidA, cyd, and lacY genes were observed only for E. coli isolates and strains.
Conclusion: PCR method proved capable to distinguish E. coli from Shigella as the most phylogenetically related genus and contrary to the classical methods, it could detect enterohaemorrhagic strains as Escherichia coli.
Keywords: Escherichia coli. Indicator microorganism, Molecular identification, Polymerase Chain Reaction
Type of Study: Original |
Subject:
Microbiology and Immunology
Received: 2018/01/13 | Accepted: 2018/04/29 | Published: 2018/11/3
Received: 2018/01/13 | Accepted: 2018/04/29 | Published: 2018/11/3
References
1. Horakova K, Mlejnkova H, Mlejnek P. Specific detection of Escherichia coli isolated from water samples using polymerase chain reaction targeting four genes: cytochrome bd complex, lactose permease, β‐d‐glucuronidase, and β‐d‐ galactosidase. J Appl Microbiol 2008; 105(4): 970-6.
2. Croxen MA, Finlay BB. Molecular mechanisms of Escherichia coli pathogenicity. Nat Rev Microbiol 2010; 8(1): 26-38.
3. Moresco V, Viancelli A, Nascimento MA, et al. Microbiological and physicochemical analysis of the coastal waters of southern Brazil. Mar Pollut Bull 2012; 64(1): 40-8.
4. Molina F, Lopez-Acedo E, Tabla R, et al. Improved detection of Escherichia coli and coliform bacteria by multiplex PCR. BMC Biotechnol 2015; 15(48): 1-9.
5. Rompre A, Servais P, Baudart J, et al. Detection and enumeration of coliforms in drinking water: current methods and emerging approaches. J Microbiol Methods 2002; 49(1): 31-54.
6. Horáková K, Mlejnková H, Mlejnek P. Direct detection of bacterial faecal indicators in water samples using PCR. Water Sci Technol 2006; 54(3): 135-40.
7. Augoustinos MT, Grabow NA, Genthe B, et al. An Improved Membrane Filtration Method for Enumeration of Faecal Coliforms and E. Coli by a Fluorogenic β-Glucuronidase Assay. Water Science and Technology 1993; 27(3-4): 267-70.
8. Monday SR, Whittam TS, Feng PC. Genetic and evolutionary analysis of mutations in the gusA gene that cause the absence of beta-glucuronidase activity in Escherichia coli O157:H7. J Infect Dis 2001; 184(7): 918-21.
9. Abbasi P, Kargar M, Doosti A, et al. Characterization of Shiga-toxin producing E. coli (STEC) and enteropathogenic E. coli (EPEC) using multiplex Real-Time PCR assays for stx1, stx2, eaeA. Iran J Microbiol 2014; 6(3): 169-74.
10. Abbasi P, Kargar M, Doosti A, et al. Molecular Detection of Diffusely Adherent Escherichia coli Strains Associated with Diarrhea in Shiraz, Iran. Arch Pediatr Infect Dis 2017; 5(2): e37629.
11. Bej AK, Steffan RJ, DiCesare J, et al. Detection of coliform bacteria in water by polymerase chain reaction and gene probes. Appl Environ Microbiol 1990; 56(2): 307-14.
12. Bej AK, McCarty SC, Atlas R. Detection of coliform bacteria and Escherichia coli by multiplex polymerase chain reaction: comparison with defined substrate and plating methods for water quality monitoring. Appl Environ Microbiol 1991; 57(8): 2429-32.
13. Løbersli I, Wester A, Kristiansen Å, et al. Molecular differentiation of Shigella spp. from enteroinvasive E. coli. Eur J Microbiol Immunol (Bp) 2016; 6(3): 197-205.
14. Dehghan F, Zolfaghari MR, Arjomandzadegan M, 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(5): 404-9.
15. Abtahi H, Ghannadzadeh M, Salmanian AH, et al. Improvement of PCR in detection of coliform in water pollution. AMUJ 2008; 11(3): 1-7.
16. Almeida C, Soares F. Microbiological monitoring of bivalves from the Ria Formosa Lagoon (south coast of Portugal): a 20 years of sanitary survey. Mar Pollut Bull 2012; 64(2): 252-62.
17. Ito H, Kido N, Arakawa Y, et al. Possible mechanisms underlying the slow lactose fermentation phenotype in Shigella spp. Appl Environ Microbiol 1991; 57(10): 2912-7.
18. Dehghan F, Zolfaghari MR, Arjomandzadegan M, et al. Comparison of PCR with Standard Method (MPN) for detection of bacterial contamination in drinking water. Iran South Med J 2014; 17(5): 867-78. (Persian)
| Rights and Permissions | |
 |
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |