Next Article in Journal
Dual-Specific Interaction to Detect DNA on Gold Nanoparticles
Next Article in Special Issue
Immunoassays and Biosensors for the Detection of Cyanobacterial Toxins in Water
Previous Article in Journal
Design and Characterization of a Fully Differential MEMS Accelerometer Fabricated Using MetalMUMPs Technology
Previous Article in Special Issue
Performance Improvement of the One-Dot Lateral Flow Immunoassay for Aflatoxin B1 by Using a Smartphone-Based Reading System
Sensors 2013, 13(5), 5737-5748; doi:10.3390/s130505737
Article

A High-Throughput Antibody-Based Microarray Typing Platform

1,* , 2
, 1
, 3
, 4
, 2
, 1
, 5
, 3
, 2
, 5
, 5
, 1
, 2
 and 1
Received: 22 February 2013; in revised form: 9 April 2013 / Accepted: 15 April 2013 / Published: 3 May 2013
View Full-Text   |   Download PDF [1535 KB, uploaded 21 June 2014]   |   Browse Figures
Abstract: Many rapid methods have been developed for screening foods for the presence of pathogenic microorganisms. Rapid methods that have the additional ability to identify microorganisms via multiplexed immunological recognition have the potential for classification or typing of microbial contaminants thus facilitating epidemiological investigations that aim to identify outbreaks and trace back the contamination to its source. This manuscript introduces a novel, high throughput typing platform that employs microarrayed multiwell plate substrates and laser-induced fluorescence of the nucleic acid intercalating dye/stain SYBR Gold for detection of antibody-captured bacteria. The aim of this study was to use this platform for comparison of different sets of antibodies raised against the same pathogens as well as demonstrate its potential effectiveness for serotyping. To that end, two sets of antibodies raised against each of the “Big Six” non-O157 Shiga toxin-producing E. coli (STEC) as well as E. coli O157:H7 were array-printed into microtiter plates, and serial dilutions of the bacteria were added and subsequently detected. Though antibody specificity was not sufficient for the development of an STEC serotyping method, the STEC antibody sets performed reasonably well exhibiting that specificity increased at lower capture antibody concentrations or, conversely, at lower bacterial target concentrations. The favorable results indicated that with sufficiently selective and ideally concentrated sets of biorecognition elements (e.g., antibodies or aptamers), this high-throughput platform can be used to rapidly type microbial isolates derived from food samples within ca. 80 min of total assay time. It can also potentially be used to detect the pathogens from food enrichments and at least serve as a platform for testing antibodies.
Keywords: antibody; microarray; bacteria; fluorescence; microtiter plate; typing antibody; microarray; bacteria; fluorescence; microtiter plate; typing
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Export to BibTeX |
EndNote


MDPI and ACS Style

Gehring, A.; Barnett, C.; Chu, T.; DebRoy, C.; D'Souza, D.; Eaker, S.; Fratamico, P.; Gillespie, B.; Hegde, N.; Jones, K.; Lin, J.; Oliver, S.; Paoli, G.; Perera, A.; Uknalis, J. A High-Throughput Antibody-Based Microarray Typing Platform. Sensors 2013, 13, 5737-5748.

AMA Style

Gehring A, Barnett C, Chu T, DebRoy C, D'Souza D, Eaker S, Fratamico P, Gillespie B, Hegde N, Jones K, Lin J, Oliver S, Paoli G, Perera A, Uknalis J. A High-Throughput Antibody-Based Microarray Typing Platform. Sensors. 2013; 13(5):5737-5748.

Chicago/Turabian Style

Gehring, Andrew; Barnett, Charles; Chu, Ted; DebRoy, Chitrita; D'Souza, Doris; Eaker, Shannon; Fratamico, Pina; Gillespie, Barbara; Hegde, Narasimha; Jones, Kevin; Lin, Jun; Oliver, Stephen; Paoli, George; Perera, Ashan; Uknalis, Joseph. 2013. "A High-Throughput Antibody-Based Microarray Typing Platform." Sensors 13, no. 5: 5737-5748.


Sensors EISSN 1424-8220 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert