{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,14]],"date-time":"2025-10-14T06:59:24Z","timestamp":1760425164405,"version":"build-2065373602"},"reference-count":15,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2013,5,3]],"date-time":"2013-05-03T00:00:00Z","timestamp":1367539200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/3.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Sensors"],"abstract":"<jats:p>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 \u201cBig Six\u201d  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.<\/jats:p>","DOI":"10.3390\/s130505737","type":"journal-article","created":{"date-parts":[[2013,5,3]],"date-time":"2013-05-03T10:55:42Z","timestamp":1367578542000},"page":"5737-5748","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["A High-Throughput Antibody-Based Microarray  Typing Platform"],"prefix":"10.3390","volume":"13","author":[{"given":"Andrew","family":"Gehring","sequence":"first","affiliation":[{"name":"Molecular Characterization of Foodborne Pathogens Research Unit, United States Department of Agriculture-North Atlantic Area-Agricultural Research Service-Eastern Regional Research Center, Wyndmoor, PA 19038, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Charles","family":"Barnett","sequence":"additional","affiliation":[{"name":"NanoDetection Technology, Inc., Franklin, OH 45005, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ted","family":"Chu","sequence":"additional","affiliation":[{"name":"Molecular Characterization of Foodborne Pathogens Research Unit, United States Department of Agriculture-North Atlantic Area-Agricultural Research Service-Eastern Regional Research Center, Wyndmoor, PA 19038, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chitrita","family":"DebRoy","sequence":"additional","affiliation":[{"name":"Reference Center, Department of Veterinary and Biomedical Sciences,  Pennsylvania State University, University Park, PA 16802, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Doris","family":"D'Souza","sequence":"additional","affiliation":[{"name":"Department of Food Science and Technology, University of Tennessee, Knoxville, TN 37916, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shannon","family":"Eaker","sequence":"additional","affiliation":[{"name":"NanoDetection Technology, Inc., Franklin, OH 45005, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pina","family":"Fratamico","sequence":"additional","affiliation":[{"name":"Molecular Characterization of Foodborne Pathogens Research Unit, United States Department of Agriculture-North Atlantic Area-Agricultural Research Service-Eastern Regional Research Center, Wyndmoor, PA 19038, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Barbara","family":"Gillespie","sequence":"additional","affiliation":[{"name":"Department of Animal Science, University of Tennessee, Knoxville, TN 37916, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Narasimha","family":"Hegde","sequence":"additional","affiliation":[{"name":"Reference Center, Department of Veterinary and Biomedical Sciences,  Pennsylvania State University, University Park, PA 16802, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kevin","family":"Jones","sequence":"additional","affiliation":[{"name":"NanoDetection Technology, Inc., Franklin, OH 45005, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jun","family":"Lin","sequence":"additional","affiliation":[{"name":"Department of Animal Science, University of Tennessee, Knoxville, TN 37916, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Stephen","family":"Oliver","sequence":"additional","affiliation":[{"name":"Department of Animal Science, University of Tennessee, Knoxville, TN 37916, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"George","family":"Paoli","sequence":"additional","affiliation":[{"name":"Molecular Characterization of Foodborne Pathogens Research Unit, United States Department of Agriculture-North Atlantic Area-Agricultural Research Service-Eastern Regional Research Center, Wyndmoor, PA 19038, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ashan","family":"Perera","sequence":"additional","affiliation":[{"name":"NanoDetection Technology, Inc., Franklin, OH 45005, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joseph","family":"Uknalis","sequence":"additional","affiliation":[{"name":"Molecular Characterization of Foodborne Pathogens Research Unit, United States Department of Agriculture-North Atlantic Area-Agricultural Research Service-Eastern Regional Research Center, Wyndmoor, PA 19038, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2013,5,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"7","DOI":"10.3201\/eid1701.P11101","article-title":"Foodborne illness acquired in the United States\u2014Major pathogens","volume":"17","author":"Scallan","year":"2011","journal-title":"Emerging Infect. Dis."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Sofos, J. (2005). Improving the Safety of Fresh Meat, Woodhead Publishing, Ltd.","DOI":"10.1201\/9781439823521"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"710","DOI":"10.1016\/j.fm.2010.04.008","article-title":"Alternative microbial methods: An overview and selection criteria","volume":"27","author":"Jasson","year":"2010","journal-title":"Food Microbiol"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1007\/s00604-012-0824-3","article-title":"Recent developments in rapid multiplexed bioanalytical methods for foodborne pathogenic bacteria detection","volume":"178","author":"Roda","year":"2012","journal-title":"Microchimica Acta"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"232","DOI":"10.1016\/j.biotechadv.2009.12.004","article-title":"An overview of foodborne pathogen detection: In the perspective of biosensors","volume":"28","author":"Velusamy","year":"2010","journal-title":"Biotechnol. Adv."},{"key":"ref_6","first-page":"225","article-title":"Bacterial typing methods","volume":"26","year":"2009","journal-title":"Acta Fac. Med. Naiss."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"5","DOI":"10.2116\/analsci.23.5","article-title":"The array biosensor: Portable, automated systems","volume":"23","author":"Ligler","year":"2007","journal-title":"Anal. Sci.: Int. J. Jpn. Soc. Anal. Chem."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1146\/annurev-anchem-061010-114010","article-title":"High-throughput biosensors for multiplexed food-borne pathogen detection","volume":"4","author":"Gehring","year":"2011","journal-title":"Annu. Rev. Anal. Chem."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1016\/j.mcp.2009.10.005","article-title":"Development of an oligonucleotide-based microarray to detect multiple foodborne pathogens","volume":"24","author":"Suo","year":"2010","journal-title":"Mol.Cell Probes"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"561","DOI":"10.1128\/CVI.13.5.561-567.2006","article-title":"Use of miniaturized protein arrays for Escherichia coli O serotyping","volume":"13","author":"Anjum","year":"2006","journal-title":"Clin. Vaccine. Immunol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1006\/abio.1998.2597","article-title":"Use of a light-addressable potentiometric sensor for the detection of Escherichia coli O157:H7","volume":"258","author":"Gehring","year":"1998","journal-title":"Anal. Biochem."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1760","DOI":"10.1126\/science.289.5485.1760","article-title":"Printing proteins as microarrays for high-throughput function determination","volume":"289","author":"MacBeath","year":"2000","journal-title":"Science"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"6601","DOI":"10.1021\/ac0608467","article-title":"Antibody microarray detection of Escherichia coli O157:H7: Quantification, assay limitations, and capture efficiency","volume":"78","author":"Gehring","year":"2006","journal-title":"Anal. Chem."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"819","DOI":"10.4315\/0362-028x.jfp-11-430","article-title":"Latex agglutination assays for detection of Non-O157 shiga toxin producing Escherichia coli Serogroups O26, O45, O103, O111, O121, and O145","volume":"75","author":"Medina","year":"2012","journal-title":"J. Food Prot."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Archer, C.T., Kim, J.F., Jeong, H., Park, J.H., Vickers, C.E., Lee, S.Y., and Nielsen, L.K. (2011). The genome sequence of E. coliW (ATCC 9637): Comparative genome analysis and an improved genome-scale reconstruction of E. coli. BMC Genomics.","DOI":"10.1186\/1471-2164-12-9"}],"container-title":["Sensors"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1424-8220\/13\/5\/5737\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T21:46:31Z","timestamp":1760219191000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1424-8220\/13\/5\/5737"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2013,5,3]]},"references-count":15,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2013,5]]}},"alternative-id":["s130505737"],"URL":"https:\/\/doi.org\/10.3390\/s130505737","relation":{},"ISSN":["1424-8220"],"issn-type":[{"type":"electronic","value":"1424-8220"}],"subject":[],"published":{"date-parts":[[2013,5,3]]}}}