Multiplex-PCR and PCR-RFLP assays to monitor water quality against pathogenic bacteria

Abd-El-Haleem, D. ; Zaki, S. ; Rushdy, Abeer ; Abd-El-Rahiem, W. ; Kheiralla, Z.H. 


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.


Other data

Issue Date 2003
Journal Journal of Environmental Monitoring 
URI http://research.asu.edu.eg/handle/123456789/155767
DOI 6
865
http://www.scopus.com/inward/record.url?eid=2-s2.0-0348049777&partnerID=MN8TOARS
5
10.1039/b307476e


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