Multiplex-PCR and PCR-RFLP assays to monitor water quality against pathogenic bacteria |
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Authors: | Abd-El-Haleem Desouky Kheiralla Zeinab H Zaki Sahar Rushdy Abeer A Abd-El-Rahiem Walaa |
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Affiliation: | Environmental Biotechnology Department, Genetic Engineering and Biotechnology Research Institute, Mubarak City for Scientific Research and Technology Applications, New Burg El-Arab City 21934, Alexandria, Egypt. abdelhaleemm@yahoo.de |
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Abstract: | In this work we developed and optimized two molecular-based approaches to monitor rapidly, sensitively and specifically bacterial pathogens from three different genera, Escherichia coli, Pseudomonas aeruginosa, and Salmonella spp., directly in waters. To achieve this aim, firstly a multiplex-PCR assay (M-PCR) was optimized using a primer pair specific for each pathogen. Secondly, as a molecular confirmatory test after isolation of the pathogens by classical microbiological methods, PCR-RFLP of their amplified 16S rDNA genes was performed. It was observed from the results that the developed M-PCR assay has significant impact on the ability to detect sensitively, rapidly and specifically the three pathogens directly in water within a short time (5 h from sampling to obtain final results), therefore it represents a considerable advancement over other known more time-consuming and less-sensitive methods for identification and characterization of these kinds of pathogens. |
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