On-chip parallel detection of foodborne pathogens using loop-mediated isothermal amplification
| العنوان: | On-chip parallel detection of foodborne pathogens using loop-mediated isothermal amplification |
|---|---|
| المؤلفون: | Carlos Armando Duarte, Bobby Reddy, Rashid Bashir, Eric Salm, Brian Dorvel |
| المصدر: | Biomedical Microdevices. 15:821-830 |
| بيانات النشر: | Springer Science and Business Media LLC, 2013. |
| سنة النشر: | 2013 |
| مصطلحات موضوعية: | DNA, Bacterial, Microinjections, Biomedical Engineering, Loop-mediated isothermal amplification, Virulence, Food Contamination, Computational biology, Biology, medicine.disease_cause, Sensitivity and Specificity, Microbiology, Listeria monocytogenes, Salmonella, Escherichia coli, medicine, Food microbiology, Molecular Biology, Polymerase, DNA Primers, Oligonucleotide Array Sequence Analysis, DNA extraction, Food Microbiology, Nucleic acid, biology.protein, Nucleic Acid Amplification Techniques |
| الوصف: | According to estimates issued by the Center for Disease Control and Prevention, one out of six Americans will get sick during this year due to consumption of contaminated products and there will be 50,000 related hospitalizations. To control and treat the responsible foodborne diseases, rapid and accurate detection of pathogens is extremely important. A portable device capable of performing nucleic acid amplification will enable the effective detection of infectious agents in multiple settings, leading to better enforcement of food safety regulations. This work demonstrates the multiplexed detection of food pathogens through loop-mediated isothermal amplification on a silicon chip. Silane passivation is used to prevent the adsorption of the polymerase on silicon oxide, which can severely inhibit nucleic acid amplification. We demonstrate the multiplexed screening of virulence genes of Listeria monocytogenes, Escherichia coli, and Salmonella by dehydrating the corresponding primers in oxidized silicon wells. Droplets of 30 nL with reagents for nucleic acid amplification and lysate of suspected pathogens are arrayed on micro-machined wells with an automated microinjection system. We show that dehydrated primers re-suspend when other reagents are microinjected, and the resulting mix can be used to specifically amplify the targeted gene. Results of characterization experiments demonstrate sensitivity down to a few templates per reaction, specificity that enables multiplexed screening, and robustness that allows amplification without DNA extraction. |
| تدمد: | 1572-8781 1387-2176 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::3e4ea3980678e358dd22d51f3adb5d08 https://doi.org/10.1007/s10544-013-9769-5 |
| حقوق: | CLOSED |
| رقم الأكسشن: | edsair.doi.dedup.....3e4ea3980678e358dd22d51f3adb5d08 |
| قاعدة البيانات: | OpenAIRE |
Find this article in full text from Springer Verlag. | تدمد: | 15728781 13872176 |
|---|