A versatile-deployable bacterial detection system for food and environmental safety based on LabTube-automated DNA purification, LabReader-integrated amplification, readout and analysis
Contamination of foods is a public health hazard that episodically causes thousands of deaths and sickens millions worldwide. To ensure food safety and quality, rapid, low-cost and easy-to-use detection methods are desirable. Here, the LabSystem is introduced for integrated, automated DNA purificati...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | en_US |
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Royal Society of Chemistry
2015
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| Online Access: | http://hdl.handle.net/1721.1/98225 https://orcid.org/0000-0002-5048-4109 |
| _version_ | 1826190750583357440 |
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| author | Hoehl, Melanie M. Bocholt, Eva Schulte Kloke, Arne Paust, Nils Stetten, Felix von Zengerle, Roland Steigert, Juergen Slocum, Alexander H. |
| author2 | Harvard University--MIT Division of Health Sciences and Technology |
| author_facet | Harvard University--MIT Division of Health Sciences and Technology Hoehl, Melanie M. Bocholt, Eva Schulte Kloke, Arne Paust, Nils Stetten, Felix von Zengerle, Roland Steigert, Juergen Slocum, Alexander H. |
| author_sort | Hoehl, Melanie M. |
| collection | MIT |
| description | Contamination of foods is a public health hazard that episodically causes thousands of deaths and sickens millions worldwide. To ensure food safety and quality, rapid, low-cost and easy-to-use detection methods are desirable. Here, the LabSystem is introduced for integrated, automated DNA purification, amplification and detection. It consists of a disposable, centrifugally driven DNA purification platform (LabTube) and a low-cost UV/vis-reader (LabReader). For demonstration of the LabSystem in the context of food safety, purification of Escherichia coli (non-pathogenic E. coli and pathogenic verotoxin-producing E. coli (VTEC)) in water and milk and the product-spoiler Alicyclobacillus acidoterrestris (A. acidoterrestris) in apple juice was integrated and optimized in the LabTube. Inside the LabReader, the purified DNA was amplified, readout and analyzed using both qualitative isothermal loop-mediated DNA amplification (LAMP) and quantitative real-time PCR. For the LAMP-LabSystem, the combined detection limits for purification and amplification of externally lysed VTEC and A. acidoterrestris are 10[superscript 2]–10[superscript 3] cell-equivalents. In the PCR-LabSystem for E. coli cells, the quantification limit is 10[superscript 2] cell-equivalents including LabTube-integrated lysis. The demonstrated LabSystem only requires a laboratory centrifuge (to operate the disposable, fully closed LabTube) and a low-cost LabReader for DNA amplification, readout and analysis. Compared with commercial DNA amplification devices, the LabReader improves sensitivity and specificity by the simultaneous readout of four wavelengths and the continuous readout during temperature cycling. The use of a detachable eluate tube as an interface affords semi-automation of the LabSystem, which does not require specialized training. It reduces the hands-on time from about 50 to 3 min with only two handling steps: sample input and transfer of the detachable detection tube. |
| first_indexed | 2024-09-23T08:45:08Z |
| format | Article |
| id | mit-1721.1/98225 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T08:45:08Z |
| publishDate | 2015 |
| publisher | Royal Society of Chemistry |
| record_format | dspace |
| spelling | mit-1721.1/982252022-09-30T10:56:01Z A versatile-deployable bacterial detection system for food and environmental safety based on LabTube-automated DNA purification, LabReader-integrated amplification, readout and analysis Hoehl, Melanie M. Bocholt, Eva Schulte Kloke, Arne Paust, Nils Stetten, Felix von Zengerle, Roland Steigert, Juergen Slocum, Alexander H. Harvard University--MIT Division of Health Sciences and Technology Massachusetts Institute of Technology. Department of Mechanical Engineering Slocum, Alexander H. Hoehl, Melanie M. Contamination of foods is a public health hazard that episodically causes thousands of deaths and sickens millions worldwide. To ensure food safety and quality, rapid, low-cost and easy-to-use detection methods are desirable. Here, the LabSystem is introduced for integrated, automated DNA purification, amplification and detection. It consists of a disposable, centrifugally driven DNA purification platform (LabTube) and a low-cost UV/vis-reader (LabReader). For demonstration of the LabSystem in the context of food safety, purification of Escherichia coli (non-pathogenic E. coli and pathogenic verotoxin-producing E. coli (VTEC)) in water and milk and the product-spoiler Alicyclobacillus acidoterrestris (A. acidoterrestris) in apple juice was integrated and optimized in the LabTube. Inside the LabReader, the purified DNA was amplified, readout and analyzed using both qualitative isothermal loop-mediated DNA amplification (LAMP) and quantitative real-time PCR. For the LAMP-LabSystem, the combined detection limits for purification and amplification of externally lysed VTEC and A. acidoterrestris are 10[superscript 2]–10[superscript 3] cell-equivalents. In the PCR-LabSystem for E. coli cells, the quantification limit is 10[superscript 2] cell-equivalents including LabTube-integrated lysis. The demonstrated LabSystem only requires a laboratory centrifuge (to operate the disposable, fully closed LabTube) and a low-cost LabReader for DNA amplification, readout and analysis. Compared with commercial DNA amplification devices, the LabReader improves sensitivity and specificity by the simultaneous readout of four wavelengths and the continuous readout during temperature cycling. The use of a detachable eluate tube as an interface affords semi-automation of the LabSystem, which does not require specialized training. It reduces the hands-on time from about 50 to 3 min with only two handling steps: sample input and transfer of the detachable detection tube. 2015-08-25T19:01:39Z 2015-08-25T19:01:39Z 2014-03 2014-01 Article http://purl.org/eprint/type/JournalArticle 0003-2654 1364-5528 http://hdl.handle.net/1721.1/98225 Hoehl, Melanie M., Eva Schulte Bocholt, Arne Kloke, Nils Paust, Felix von Stetten, Roland Zengerle, Juergen Steigert, and Alexander H. Slocum. “A Versatile-Deployable Bacterial Detection System for Food and Environmental Safety Based on LabTube-Automated DNA Purification, LabReader-Integrated Amplification, Readout and Analysis.” Analyst 139, no. 11 (2014): 2788. https://orcid.org/0000-0002-5048-4109 en_US http://dx.doi.org/10.1039/c4an00123k The Analyst Creative Commons Attribution-Noncommercial-Share Alike http://creativecommons.org/licenses/by-nc-sa/4.0/ application/pdf Royal Society of Chemistry Prof. Slocum via Angie Locknar |
| spellingShingle | Hoehl, Melanie M. Bocholt, Eva Schulte Kloke, Arne Paust, Nils Stetten, Felix von Zengerle, Roland Steigert, Juergen Slocum, Alexander H. A versatile-deployable bacterial detection system for food and environmental safety based on LabTube-automated DNA purification, LabReader-integrated amplification, readout and analysis |
| title | A versatile-deployable bacterial detection system for food and environmental safety based on LabTube-automated DNA purification, LabReader-integrated amplification, readout and analysis |
| title_full | A versatile-deployable bacterial detection system for food and environmental safety based on LabTube-automated DNA purification, LabReader-integrated amplification, readout and analysis |
| title_fullStr | A versatile-deployable bacterial detection system for food and environmental safety based on LabTube-automated DNA purification, LabReader-integrated amplification, readout and analysis |
| title_full_unstemmed | A versatile-deployable bacterial detection system for food and environmental safety based on LabTube-automated DNA purification, LabReader-integrated amplification, readout and analysis |
| title_short | A versatile-deployable bacterial detection system for food and environmental safety based on LabTube-automated DNA purification, LabReader-integrated amplification, readout and analysis |
| title_sort | versatile deployable bacterial detection system for food and environmental safety based on labtube automated dna purification labreader integrated amplification readout and analysis |
| url | http://hdl.handle.net/1721.1/98225 https://orcid.org/0000-0002-5048-4109 |
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