期刊
ANALYTICAL CHEMISTRY
卷 93, 期 9, 页码 4149-4153出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.0c05355
关键词
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资金
- Chemical Measurement and Imaging Program at the National Science Foundation [CHE-1709372]
- Alice Hudson Professorship
The research utilizes molecular beacon (MB) probes for sequence-specific detection of LAMP on LFIA strips, showing detection capability for various DNA sequences and single-nucleotide polymorphisms. Coupling the method with solid-phase microextraction allows for detection of target sequences in different samples, and a 3D printed device is designed to prevent contamination.
Loop-mediated isothermal amplification (LAMP) holds great potential for point-of-care (POC) diagnostics due to its speed and sensitivity. However, differentiation between spurious amplification and amplification of the target sequence is a challenge. Herein, we develop the use of molecular beacon (MB) probes for the sequence-specific detection of LAMP on commercially available lateral flow immunoassay (LFIA) strips. The detection of three unique DNA sequences, including ORF1a from SARS-CoV-2, is demonstrated. In addition, the method is capable of detecting clinically relevant single-nucleotide polymorphisms (BRAF V600E). For all sequences tested, the LFIA method offers similar sensitivity to fluorescence detection using a qPCR instrument. We also demonstrate the coupling of the method with solid-phase microextraction to enable isolation and detection of the target sequences from human plasma, pond water, and artificial saliva. Lastly, a 3D printed device is designed and implemented to prevent contamination caused by opening the reaction containers after LAMP.
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