Multiplex-PCR and PCR-RFLP assays to monitor water quality against pathogenic bacteria.
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.
Abd-El-Haleem
Desouky
D
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
Kheiralla
Zeinab H
ZH
Zaki
Sahar
S
Rushdy
Abeer A
AA
Abd-El-Rahiem
Walaa
W
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
D004269:DNA, Bacterial