Application of loop-mediated isothermal amplification (lamp) in microbiological water quality analysis

Abstract

Water is life thus access to safe drinking water is essential to health and a basic human right. Kenya is considered a water scarce country whose water sources are becoming more contaminated through changes in land use and poor solid waste management. This has in turn affected the quantity and quality of portable water available. There are nearly 1.7 billion cases of waterborne diarrheal disease every year with over 760 000 deaths of children under five years. Traditional methods of identification of waterborne pathogens are less specific, sensitive, timeconsuming and laborious, so there is a need for the development of innovative methods for their rapid identification. Recent advances in molecular cloning and recombinant DNA techniques have revolutionized the detection of pathogens in foods. In this study the application of a LAMP based technique for the rapid identification of the Escherichia coli, Enterococcus feacalis and Clostridia perfringens DNAs was undertaken. Suitable primers were designed based on specific gene MalB of Escherichia coli, tuf gene of Enterococcus feacalis and CPE gene of Clostridia perfringens for amplification. Selective media for the various indicator microbes were used and specific primers for both LAMP and PCR designed. The study reveals that bottled water from Majesty and Starpop companies were contaminated by coliforms while the ground water from Kikuyu Springs, rainwater from Lang’ata and tapped water from Naivasha’s Fishermans campsite were contaminated by the three target bacteria. This was possible via culture work as LAMP and PCR assays were not fully optimized to enable the detection of Escherichia coli, Enterococcus feacalis and Clostridiun perfrigens DNAs. The study shows LAMP as a viable tool in microbial water quality analysis from the little amplification achieved with the positive controls that had high DNA concentrations