Abstract
Consumption of contaminated drinking water heavily contributes to the burden of gastrointestinal waterborne diseases. Conventional detection methodologies present several shortcomings, such as indirect measure of indicator species, low throughput and time consume. DNA chips have the potential to serve as surveillance systems for the simultaneous detection of pathogens overcoming these limitations. We have developed a DNA chip for simultaneous detection of multiple waterborne pathogens. Species specific DNA probes were implemented on a microarray, for microorganism detection. Present study reports the results of one untreated water sample analyzed by conventional methods and by the DNA chip. The results were concordant for the mandatory organisms (total coliforms, <em>Escherichia coli</em>, fecal enterococci) using both methods, reinforcing the utility and proof-ofconcept of the DNA chip. However, it is necessary a prior enrichment in a culture medium in order to obtain a positive signal using the DNA chip. The DNA chip may be a valuable distinctive tool for waterborne pathogens detection
Highlights
Ciências, Centre for Biodiversity, Functional and Integrative Genomics (BioFIG), Campus da FCUL, Campo growth) and may detect stressed bacteria
It is necesstruction of a DNA chip for simultaneous detections of microorganisms in water samples[3,5] and its suitability in detecting genomic DNA isolated from a pure culture of Escherichia coli, one of the European Community mandatory indicator microorganism.[5]
We have reported the suitability of the DNA chip for detecting genomic DNA of a pure culture of E. coli.[5]
Summary
Conventional culture methods were used to determine the microbiological quality of water. Two selective medium at an appropriate temperayear could be avoided if these measures were liters of the sample were used for culture ture: i) for detection of total coliform bacteria implemented.[1] Water quality control is essential method detection and one liter for DNA chip the membrane was incubated in m-lauryl sulto assure the delivery of safe human consump- analysis. The use of a previous gy, and to reduce hybridization artefacts, as colonies tested for absence of oxidase activity; step culture was made to test if the bacterial suggested by others.[6] ii) for detection of fecal coliform bacteria, the load in the water sample is sufficient to be membrane was incubated in MLSA medium at detected by the DNA chip, or if its growth in an Image acquisition and data analysis. The high number of replicates of each probe (eight) and the semi-random distribution of the probes in each grid[5] are used to control for the high variability inherent to the methodolo-
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