LOW-COST BACTERIAL DETECTION SYSTEM FOR FOOD SAFETY BASED ON AUTOMATED DNA EXTRACTION, AMPLIFICATION AND READOUT
To ensure food, medical and environmental safety and quality, rapid, low-cost and easy-to-use detection methods are desirable. Here, the LabSystem is introduced for integrated, automated DNA purification and amplification. It consists of a disposable, centrifugally-driven DNA purification platform (...
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| Format: | Article |
| Language: | en_US |
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Chemical and Biological Microsystems Society
2013
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| Online Access: | http://hdl.handle.net/1721.1/82905 https://orcid.org/0000-0002-5048-4109 |
| _version_ | 1826196465548001280 |
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| author | Hoehl, Melanie Margarete Bocholt, Eva Schulte Karippai, Nobu 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 Margarete Bocholt, Eva Schulte Karippai, Nobu Zengerle, Roland Steigert, Juergen Slocum, Alexander H. |
| author_sort | Hoehl, Melanie Margarete |
| collection | MIT |
| description | To ensure food, medical and environmental safety and quality, rapid, low-cost and easy-to-use detection methods are desirable. Here, the LabSystem is introduced for integrated, automated DNA purification and amplification. It consists of a disposable, centrifugally-driven DNA purification platform (LabTube) and the subsequent amplification in a low-cost UV/vis-reader (LabReader). In this paper, food safety was chosen as the first sample application with pathogenic verotoxin-producing (VTEC) Escherichia coli (E.coli) in water and milk, and the product-spoiler Alicyclobacillus acidoterrestris (A acidoterrestris) in apple juice as sample organisms. DNA was amplified qualitatively using isothermal loop-mediated DNA amplification (LAMP) and quantitatively using real-time PCR. By optimizing manual purification protocols inside the LabTube, as little as 45 inserted DNA copies were extracted from E.coli and A.acidoterrestris lysates in real samples (milk, juice and water). To run isothermal DNA amplification (LAMP) and PCR inside the LabReader, temperature control as well as data analysis methods were implemented. Combined detection limits for DNA purification and amplification from bacteria lysates in real samples at 102-103 inserted copies were achieved. The demonstrated LabSystem runs with standard laboratory equipment and reduces hands-on times from 30min to 3min. |
| first_indexed | 2024-09-23T10:26:58Z |
| format | Article |
| id | mit-1721.1/82905 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T10:26:58Z |
| publishDate | 2013 |
| publisher | Chemical and Biological Microsystems Society |
| record_format | dspace |
| spelling | mit-1721.1/829052022-09-30T21:13:11Z LOW-COST BACTERIAL DETECTION SYSTEM FOR FOOD SAFETY BASED ON AUTOMATED DNA EXTRACTION, AMPLIFICATION AND READOUT Hoehl, Melanie Margarete Bocholt, Eva Schulte Karippai, Nobu Zengerle, Roland Steigert, Juergen Slocum, Alexander H. Harvard University--MIT Division of Health Sciences and Technology Massachusetts Institute of Technology. Department of Mechanical Engineering Hoehl, Melanie Margarete Hoehl, Melanie Margarete Slocum, Alexander H. To ensure food, medical and environmental safety and quality, rapid, low-cost and easy-to-use detection methods are desirable. Here, the LabSystem is introduced for integrated, automated DNA purification and amplification. It consists of a disposable, centrifugally-driven DNA purification platform (LabTube) and the subsequent amplification in a low-cost UV/vis-reader (LabReader). In this paper, food safety was chosen as the first sample application with pathogenic verotoxin-producing (VTEC) Escherichia coli (E.coli) in water and milk, and the product-spoiler Alicyclobacillus acidoterrestris (A acidoterrestris) in apple juice as sample organisms. DNA was amplified qualitatively using isothermal loop-mediated DNA amplification (LAMP) and quantitatively using real-time PCR. By optimizing manual purification protocols inside the LabTube, as little as 45 inserted DNA copies were extracted from E.coli and A.acidoterrestris lysates in real samples (milk, juice and water). To run isothermal DNA amplification (LAMP) and PCR inside the LabReader, temperature control as well as data analysis methods were implemented. Combined detection limits for DNA purification and amplification from bacteria lysates in real samples at 102-103 inserted copies were achieved. The demonstrated LabSystem runs with standard laboratory equipment and reduces hands-on times from 30min to 3min. 2013-12-10T19:44:58Z 2013-12-10T19:44:58Z 2013-10 Article http://purl.org/eprint/type/ConferenceItem http://hdl.handle.net/1721.1/82905 Hoehl, Melanie, Eva Schulte Bocholt, Nobu Karippai, Roland Zengerle, Juergen Steigert, and Alexander Slocum. "LOW-COST BACTERIAL DETECTION SYSTEM FOR FOOD SAFETY BASED ON AUTOMATED DNA EXTRACTION, AMPLIFICATION AND READOUT." In 17th International Conference on Miniaturized Systems for Chemistry and Life Sciences (µTAS 2013), Messe Freiburg in Freiburg, GERMANY, 27-31 October 2013. https://orcid.org/0000-0002-5048-4109 en_US http://www.microtas2013.org/program/MicroTAS2013_TechnicalProgram.pdf Proceedings of the 17th International Conference on Miniaturized Systems for chemistry and Life Sciences (MicroTAS 2013) Creative Commons Attribution-Noncommercial-Share Alike 3.0 http://creativecommons.org/licenses/by-nc-sa/3.0/ application/pdf Chemical and Biological Microsystems Society Melanie Hoehl |
| spellingShingle | Hoehl, Melanie Margarete Bocholt, Eva Schulte Karippai, Nobu Zengerle, Roland Steigert, Juergen Slocum, Alexander H. LOW-COST BACTERIAL DETECTION SYSTEM FOR FOOD SAFETY BASED ON AUTOMATED DNA EXTRACTION, AMPLIFICATION AND READOUT |
| title | LOW-COST BACTERIAL DETECTION SYSTEM FOR FOOD SAFETY BASED ON AUTOMATED DNA EXTRACTION, AMPLIFICATION AND READOUT |
| title_full | LOW-COST BACTERIAL DETECTION SYSTEM FOR FOOD SAFETY BASED ON AUTOMATED DNA EXTRACTION, AMPLIFICATION AND READOUT |
| title_fullStr | LOW-COST BACTERIAL DETECTION SYSTEM FOR FOOD SAFETY BASED ON AUTOMATED DNA EXTRACTION, AMPLIFICATION AND READOUT |
| title_full_unstemmed | LOW-COST BACTERIAL DETECTION SYSTEM FOR FOOD SAFETY BASED ON AUTOMATED DNA EXTRACTION, AMPLIFICATION AND READOUT |
| title_short | LOW-COST BACTERIAL DETECTION SYSTEM FOR FOOD SAFETY BASED ON AUTOMATED DNA EXTRACTION, AMPLIFICATION AND READOUT |
| title_sort | low cost bacterial detection system for food safety based on automated dna extraction amplification and readout |
| url | http://hdl.handle.net/1721.1/82905 https://orcid.org/0000-0002-5048-4109 |
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