Evaluation of Molecular Methods to Be Used in Sensors for The Detection of Bacteria in Water

Abstract

Waterborne E. coli and E. faecalis remain a major economic problem worldwide because of their significant impact on health and disease. There is a constant need for new diagnostic tools that can detect low bacterial concentrations in a more cost-and timeeffective manner. The development of rapid and simple molecular detection in situ is required where specialized laboratory services are limited. Hereby, the recovery of DNA from artificially contaminated water samples of three different DNA extraction methods was investigated. Two commercial kits (DNeasy Ultraclean Microbial Kit, Dynabeads™ DNA DIRECT™ Universal Kit) and a boiling method were evaluated. All methods produced DNA in sufficient concentration ranging 64.55-184.7 ng/μL for E. coli and E. faecalis, respectively. Then, PCR methods were applied to confirm the effectiveness of the process. All amplifications had an LOD ranging from 102 to 101. Here is suggested that boiling extraction could be used as a method with rapid results, easy in use, adequate efficiency, which could be included in a portable sensor tool, as biosensor. The results of the present study highlight the importance of DNA extraction suitable for the application in a biosensor for a real time monitoring of the water samples.

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“…They are considered to be appropriate because they can be used in portable or in situ autonomous systems. Their principle is that they use a biorecognition element such as tissue, antibody, or a whole microorganism that in contact with a transducer transmits the information into a signal [19]. Biosensors will guarantee consistent and precise microbial hazard control to evaluate seawater state.…”

Automated Biosensor for Rapid Field Detection of Fecal Biomarkers in Coastal Areas

“…They are considered to be appropriate because they can be used in portable or in situ autonomous systems. Their principle is that they use a biorecognition element such as tissue, antibody, or a whole microorganism that in contact with a transducer transmits the information into a signal [19]. Biosensors will guarantee consistent and precise microbial hazard control to evaluate seawater state.…”