Journal
LAB ON A CHIP
Volume 16, Issue 3, Pages 611-621Publisher
ROYAL SOC CHEMISTRY
DOI: 10.1039/c5lc01388g
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Funding
- Major International Joint Research Program of China [11120101002]
- International Science AMP
- Technology Cooperation Program of China [2013DFG02930]
- National Instrumentation Program [2013YQ190467]
- Ministry of Higher Education (MOHE), Government of Malaysia under the High Impact Research [UM.C/HIR/MOHE/ENG/44]
- National Natural Science Foundation of China [81302029]
- Natural Science Foundation of Shaanxi Province of China [2014JQ4149]
- Fundamental Research Funds for the Central Universities in Xi'an Jiaotong University [xjj2015086]
- China Postdoctoral Science Foundation [2015 M570841]
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With advances in point-of-care testing (POCT), lateral flow assays (LFAs) have been explored for nucleic acid detection. However, biological samples generally contain complex compositions and low amounts of target nucleic acids, and currently require laborious off-chip nucleic acid extraction and amplification processes (e.g., tube-based extraction and polymerase chain reaction (PCR)) prior to detection. To the best of our knowledge, even though the integration of DNA extraction and amplification into a paper-based biosensor has been reported, a combination of LFA with the aforementioned steps for simple colorimetric readout has not yet been demonstrated. Here, we demonstrate for the first time an integrated paper-based biosensor incorporating nucleic acid extraction, amplification and visual detection or quantification using a smartphone. A handheld battery-powered heating device was specially developed for nucleic acid amplification in POC settings, which is coupled with this simple assay for rapid target detection. The biosensor can successfully detect Escherichia coli (as a model analyte) in spiked drinking water, milk, blood, and spinach with a detection limit of as low as 10-1000 CFU mL(-1), and Streptococcus pneumonia in clinical blood samples, highlighting its potential use in medical diagnostics, food safety analysis and environmental monitoring. As compared to the lengthy conventional assay, which requires more than 5 hours for the entire sample-to-answer process, it takes about 1 hour for our integrated biosensor. The integrated biosensor holds great potential for detection of various target analytes for wide applications in the near future.
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