Title: Dark field microscopy-based biosensor for the detection of E. Coli in environmental water samples
Authors: LA SPINA RITAANTONIO DIANA C.DESMET CLOEVALSESIA ANDREABOMBERA RADOSLAW WIKTORNORLEN HEDVIGLETTIERI TERESACOLPO PASCAL
Citation: SENSORS vol. 19 no. 21 p. 4652
Publisher: MDPI
Publication Year: 2019
JRC N°: JRC118107
ISSN: 1424-8220 (online)
URI: https://www.mdpi.com/1424-8220/19/21/4652
https://publications.jrc.ec.europa.eu/repository/handle/JRC118107
DOI: 10.3390/s19214652
Type: Articles in periodicals and books
Abstract: Development of sensitive methods for the determination of E. coli bacteria contamination in water distribution systems is of paramount importance to ensure the microbial safety of drinking water. This work presents a new sensing platform enabling the fast detection of bacteria in field samples by using specific antibodies as the biorecognition element and dark field microscopy as the detection technique. The development of the sensing platform was performed using non-pathogenic bacteria, with the E. coli DH5 strain as the target, and Bacillus sp. 9727 as the negative control. The identification of the captured bacteria was made by analyzing the dark field microscopy images and screening the detected objects by using object circularity and size parameters. Specificity tests revealed the low unspecific attachment of either E. coli over human serum albumin antibodies (negative control for antibody specificity) and of Bacillus sp. over E. coli antibodies. The system performance was tested using field samples, collected from a wastewater treatment plant, and compared with two quantification techniques (i.e., Colilert-18 test and quantitative polymerase chain reaction (qPCR)). The results showed comparable quantification capability. Nevertheless, the present method has the advantage of being faster, is easily adaptable to in-field analysis, and can potentially be extended to the detection of other bacterial strains.
JRC Directorate:Sustainable Resources

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