Low-cost microarray platform to detect antibiotic resistance genes
Versatile DNA-based approaches for the detection of pathogens and antibiotic resistance genes have been introduced to facilitate faster diagnostic test. This requires a notable amount of cost-intensive commercially labelled DNA oligonucleotides in high-throughput assays. Here, we introduce an altern...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2019-04-01
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| Series: | Sensing and Bio-Sensing Research |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2214180418301132 |
| _version_ | 1819077904806445056 |
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| author | Noa Wolff Michaela Hendling Silvia Schönthaler Andreas F. Geiss Ivan Barišić |
| author_facet | Noa Wolff Michaela Hendling Silvia Schönthaler Andreas F. Geiss Ivan Barišić |
| author_sort | Noa Wolff |
| collection | DOAJ |
| description | Versatile DNA-based approaches for the detection of pathogens and antibiotic resistance genes have been introduced to facilitate faster diagnostic test. This requires a notable amount of cost-intensive commercially labelled DNA oligonucleotides in high-throughput assays. Here, we introduce an alternative method that utilizes a terminal deoxynucleotidyl transferase reaction to significantly reduce expenses of oligonucleotides. Antibiotic resistance genes that are of high clinical relevance were used as relevant application. The alternative labelling was based on an elongation step in the presence of biotin-conjugated nucleotides. Fluorescence-labelled streptavidin was bound to biotin-conjugated nucleotides and detected via a standard high-throughput microarray scanner. The self-labelled detection oligonucleotides performed equally well regarding sensitivity and specificity compared to commercially labelled ones, but at only 10% of their cost. Keywords: Antibiotic resistance, High-throughput detection, DNA microarray, Multiplex detection, Transferase reaction |
| first_indexed | 2024-12-21T19:04:37Z |
| format | Article |
| id | doaj.art-476d3278ff5a430fa3aac3f35cf58e9d |
| institution | Directory Open Access Journal |
| issn | 2214-1804 |
| language | English |
| last_indexed | 2024-12-21T19:04:37Z |
| publishDate | 2019-04-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Sensing and Bio-Sensing Research |
| spelling | doaj.art-476d3278ff5a430fa3aac3f35cf58e9d2022-12-21T18:53:23ZengElsevierSensing and Bio-Sensing Research2214-18042019-04-0123Low-cost microarray platform to detect antibiotic resistance genesNoa Wolff0Michaela Hendling1Silvia Schönthaler2Andreas F. Geiss3Ivan Barišić4Molecular Diagnostics, AIT Austrian Institute of Technology, Giefinggasse 4, Vienna 1210, AustriaMolecular Diagnostics, AIT Austrian Institute of Technology, Giefinggasse 4, Vienna 1210, AustriaMolecular Diagnostics, AIT Austrian Institute of Technology, Giefinggasse 4, Vienna 1210, AustriaUniversity of Natural Resources and Life Sciences, Vienna, AustriaMolecular Diagnostics, AIT Austrian Institute of Technology, Giefinggasse 4, Vienna 1210, Austria; Corresponding author.Versatile DNA-based approaches for the detection of pathogens and antibiotic resistance genes have been introduced to facilitate faster diagnostic test. This requires a notable amount of cost-intensive commercially labelled DNA oligonucleotides in high-throughput assays. Here, we introduce an alternative method that utilizes a terminal deoxynucleotidyl transferase reaction to significantly reduce expenses of oligonucleotides. Antibiotic resistance genes that are of high clinical relevance were used as relevant application. The alternative labelling was based on an elongation step in the presence of biotin-conjugated nucleotides. Fluorescence-labelled streptavidin was bound to biotin-conjugated nucleotides and detected via a standard high-throughput microarray scanner. The self-labelled detection oligonucleotides performed equally well regarding sensitivity and specificity compared to commercially labelled ones, but at only 10% of their cost. Keywords: Antibiotic resistance, High-throughput detection, DNA microarray, Multiplex detection, Transferase reactionhttp://www.sciencedirect.com/science/article/pii/S2214180418301132 |
| spellingShingle | Noa Wolff Michaela Hendling Silvia Schönthaler Andreas F. Geiss Ivan Barišić Low-cost microarray platform to detect antibiotic resistance genes Sensing and Bio-Sensing Research |
| title | Low-cost microarray platform to detect antibiotic resistance genes |
| title_full | Low-cost microarray platform to detect antibiotic resistance genes |
| title_fullStr | Low-cost microarray platform to detect antibiotic resistance genes |
| title_full_unstemmed | Low-cost microarray platform to detect antibiotic resistance genes |
| title_short | Low-cost microarray platform to detect antibiotic resistance genes |
| title_sort | low cost microarray platform to detect antibiotic resistance genes |
| url | http://www.sciencedirect.com/science/article/pii/S2214180418301132 |
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