Development of an integrated method of concentration and immunodetection of bacteria
Por:
Ezenarro, JJ, Uria, N, Castillo-Fernandez, O, Parraga, N, Sabria, M and Pascual, FXM
Publicada:
1 ene 2018
Resumen:
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.
Filiaciones:
Ezenarro, JJ:
Esfera UAB CEI, Inst Microelect Barcelona, CNM CSIC, Campus UAB, Bellaterra 08193, Spain
Uria, N:
Esfera UAB CEI, Inst Microelect Barcelona, CNM CSIC, Campus UAB, Bellaterra 08193, Spain
Castillo-Fernandez, O:
Esfera UAB CEI, Inst Microelect Barcelona, CNM CSIC, Campus UAB, Bellaterra 08193, Spain
:
Fundacio Inst Invest Germans Trias & Pujol, Unitat Malalties Infeccioses, Badalona 08916, Spain
CIBER Enfermedades Resp, Madrid 28029, Spain
Sabria, M:
CIBER Enfermedades Resp, Madrid 28029, Spain
Univ Autonoma Barcelona, Cerdanyola Del Valles 08193, Spain
Pascual, FXM:
Esfera UAB CEI, Inst Microelect Barcelona, CNM CSIC, Campus UAB, Bellaterra 08193, Spain
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