title: Multiplex-PCR and PCR-RFLP assays to monitor water quality against pathogenic bacteria.
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.
authors.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
authors.forename: Desouky
authors.initials: D
authors.lastname: Abd-El-Haleem
authors.forename: Zeinab H
authors.initials: ZH
authors.lastname: Kheiralla
authors.forename: Sahar
authors.initials: S
authors.lastname: Zaki
authors.forename: Abeer A
authors.initials: AA
authors.lastname: Rushdy
authors.forename: Walaa
authors.initials: W
authors.lastname: Abd-El-Rahiem
mesh_terms: D004269:DNA, Bacterial / Q000032:analysis; D004926:Escherichia coli / Q000235:genetics* / Q000472:pathogenicity; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D011550:Pseudomonas aeruginosa / Q000235:genetics* / Q000472:pathogenicity; D012475:Salmonella / Q000235:genetics* / Q000472:pathogenicity; D012680:Sensitivity and Specificity; D014871:Water Microbiology
chemical_list: D004269:DNA, Bacterial