Development and Validation of Multiplex PCR for the Simultaneous Detection of Total Coliform Bacteria, Escherichia coli and Clostridium perfringens in Water

Hooshyar Akbari  Khalilabad; Fatemeh  Rezaei; Elham  Khosravi; Kiarash  Ghazvini

doi:10.18502/jmb.v13i3.19508

Hooshyar Akbari Khalilabad Department of Microbiology and Virology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
Fatemeh Rezaei Department of Microbiology and Virology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
Elham Khosravi Department of Microbiology and Virology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
Kiarash Ghazvini Department of Microbiology and Virology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.

DOI: https://doi.org/10.18502/jmb.v13i3.19508

Keywords: Clostridium perfringens, E. coli, Multiplex PCR, Coliforms, Total coliform, Water

Abstract

Background: Water is an important source of numerous infectious diseases in humans. The most important indicator bacteria include Escherichia coli and total coliform species. Clostridium perfringens is a general indicator of treatment efficiency.

Methods: Diagnosing microbial contamination of water by culture method, in addition to high cost, has low speed and many limitations. The PCR (Polymerase Chain Reaction) method is a promising alternative for simultaneous detection of these bacteria. In this research, we developed multiplex PCR for detection of lacZ, uidA and plc genes as an indicator of the presence of total coliforms, Escherichia coli and Clostridium perfringens. In the following, 33 samples of water and wastewater from Razavi Khorasan province were tested using standard culture methods and this multiplex PCR for comparison.

Results: Multiplex PCR was not only confirmed by cultural results but also has more accuracy and sharpness. The results of the validation tests showed that our multiplex PCR method had a significant advantage over the conventional culture method.

Conclusion: The developed multiplex PCR method demonstrated superior accuracy and efficiency compared to traditional culture techniques, offering a reliable alternative for rapid detection of waterborne pathogens