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A digital microfluidic system for loop-mediated isothermal amplification and sequence specific pathogen detection
Liang Wan1,2; Tianlan Chen1; Jie Gao1; Cheng Dong1,2; Ada Hang-Heng Wong3; Yanwei Jia1; Pui-In Mak1,2; Chu-Xia Deng3; Rui P. Martins1,2,4
2017-11-06
Source PublicationSCIENTIFIC REPORTS
ISSN2045-2322
Volume7Pages:14586
Abstract

A digital microfluidic (DMF) system has been developed for loop-mediated isothermal amplification (LAMP)-based pathogen nucleic acid detection using specific low melting temperature (T-m ) Molecular Beacon DNA probes. A positive-temperature-coefficient heater with a temperature sensor for real-time thermal regulation was integrated into the control unit, which generated actuation signals for droplet manipulation. To enhance the specificity of the LAMP reaction, low-T-m Molecular Beacon probes were designed within the single-stranded loop structures on the LAMP reaction products. In the experiments, only 1 mu L of LAMP reaction samples containing purified Trypanosoma brucei DNA were required, which represented over a 10x reduction of reagent consumption when comparing with the conventional off-chip LAMP. On-chip LAMP for unknown sample detection could be accomplished in 40 min with a detection limit of 10 copies/reaction. Also, we accomplished an on-chip melting curve analysis of the Molecular Beacon probe from 30 to 75 degrees C within 5 min, which was 3x faster than using a commercial qPCR machine. Discrimination of non-specific amplification and lower risk of aerosol contamination for on-chip LAMP also highlight the potential utilization of this system in clinical applications. The entire platform is open for further integration with sample preparation and fluorescence detection towards a total-micro-analysis system.

DOIhttp://doi.org/10.1038/s41598-017-14698-x
URLView the original
Indexed BySCI
Language英语
WOS Research AreaScience & Technology - Other Topics
WOS SubjectMultidisciplinary Sciences
WOS IDWOS:000414415000086
PublisherNATURE PUBLISHING GROUP
The Source to ArticleWOS
Fulltext Access
Citation statistics
Cited Times [WOS]:6   [WOS Record]     [Related Records in WOS]
Document TypeJournal article
CollectionINSTITUTE OF MICROELECTRONICS
Faculty of Health Sciences
Corresponding AuthorYanwei Jia
Affiliation1.State-Key Laboratory of Analog and Mixed-Signal VLSI, University of Macau, Macao, China.
2.Department of ECE, Faculty of Science and Technology, University of Macau, Macao, China.
3.Cancer Centre, Faculty of Health Sciences, University of Macau, Macao, China
4.Instituto Superior Técnico, Universidade de Lisboa, Lisboa, Portugal. Correspondence and requests for materials should be addressed to Y.J
First Author AffilicationUniversity of Macau;  Faculty of Science and Technology
Corresponding Author AffilicationUniversity of Macau
Recommended Citation
GB/T 7714
Liang Wan,Tianlan Chen,Jie Gao,et al. A digital microfluidic system for loop-mediated isothermal amplification and sequence specific pathogen detection[J]. SCIENTIFIC REPORTS,2017,7:14586.
APA Liang Wan.,Tianlan Chen.,Jie Gao.,Cheng Dong.,Ada Hang-Heng Wong.,...&Rui P. Martins.(2017).A digital microfluidic system for loop-mediated isothermal amplification and sequence specific pathogen detection.SCIENTIFIC REPORTS,7,14586.
MLA Liang Wan,et al."A digital microfluidic system for loop-mediated isothermal amplification and sequence specific pathogen detection".SCIENTIFIC REPORTS 7(2017):14586.
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