Characterization, validation and application of a DNA microarray for the detection of mandatory and other waterborne pathogens.
Culture methods for the detection of indicator bacteria are currently used for detection of waterborne bacteria. The need for an increased range of analyzed bacteria coupled with the obtainment of rapid and early results justify the development of a DNA microarray for the identification of waterborne pathogens. This DNA microarray has 16 implanted probes with a median size of 147 bases, targeting 12 different parameters, including all mandatory indicator microorganisms, such as Escherichia coli, Clostridium perfringens, Pseudomonas aeruginosa, Staphylococcus aureus, total and fecal coliforms and enterococci. The validation performed with DNA extracted from pure microbial cultures showed the suitability of the probes for detection of the target microorganism. To overcome the high dilution of water samples it was included either a prior culture step of bacterial contaminants retained after filtering 100 ml of water, or a 10-fold increase in the volume of filtered water, that resulted in the increase of the detected bacteria. The analysis of complex environmental water samples using culture methods and the DNA microarray revealed that the latter detected the same parameters plus other bacteria tested only in the DNA microarray. The results show that this DNA microarray may be a useful tool for water microbiological surveillance.
['Gomes|Maria|M|', 'Vieira|Helena|H|', 'Vale|Filipa F|FF|']
D003016:Clostridium perfringens D019295:Computational Biology D004269:DNA, Bacterial D004784:Environmental Monitoring D004926:Escherichia coli D005103:Expert Systems D005374:Filtration D007390:International Agencies D057230:Limit of Detection D058889:Molecular Typing D020411:Oligonucleotide Array Sequence Analysis D011174:Portugal D011550:Pseudomonas aeruginosa D015203:Reproducibility of Results D045483:Rivers D013211:Staphylococcus aureus D014871:Water Microbiology D060986:Water Wells D000069578:Waterborne Diseases
DNA microarray waterborne pathogen indicator bacteria bacterial detection drinking water quality