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Analytical and Bioanalytical Chemistry
Article . 2017 . Peer-reviewed
License: Springer TDM
Data sources: Crossref
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Development of an integrated method of concentration and immunodetection of bacteria

Authors: Josune J. Ezenarro; Naroa Uria; Óscar Castillo-Fernández; Noemí Párraga; Miquel Sabrià; Francesc Xavier Muñoz Pascual;

Development of an integrated method of concentration and immunodetection of bacteria

Abstract

The microbial quality of water is a key aspect to avoid environmental and public health problems. The low pathogen concentration needed to produce a disease outbreak makes it essential to process large water volumes and use sensitive and specific methods such as immunoassays for its detection. In the present work, we describe the development of a device based on microfiltration membranes to integrate the concentration and the immunodetection of waterborne bacteria. A microfiltration membrane treatment protocol was designed to reduce the non-specific binding of antibodies, for which different blocking agents were tested. Thus, the proof of concept of the microbial detection system was also carried out using Escherichia coli as the bacterial pathogen model. E. coli suspensions were filtered through the membranes at 0.5 mL s-1, and the E. coli concentration measurements were made by absorbance, at 620 nm, of the resultant product of the enzymatic reaction among the horseradish peroxidase (HRP) bonded to the antibody, and the substrate 3,3',5,5'-tetramethylbenzidine (TMB). The results showed that the homemade concentration system together with the developed membrane treatment protocol is able to detect E. coli cells with a limit of detection (LoD) of about 100 CFU in 100 mL. Graphical abstract Scheme of the integrated method of concentration and immunodetection of bacteria.

Keywords

Immunoassay, Benzidines, E. coli, Pathogen detection, Equipment Design, Sample concentration, Antibodies, Immunoenzyme Techniques, Limit of Detection, Microfiltration membrane, Escherichia coli, Humans, Water Microbiology, Escherichia coli Infections, Filtration, Horseradish Peroxidase

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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
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