A Portable Digital Loop-Mediated Isothermal Amplification Platform Based on Microgel Array and Hand-Held Reader
| Autor: | Chunyan Yao, Feng Xu, Zedong Li, Lei Cao, Minli You, Jie Hu, Xiaojin Guo, Yulin Ren, Ping Mao, Fei Li, Miao Tian |
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| Rok vydání: | 2021 |
| Předmět: |
Fluid Flow and Transfer Processes
Lung Neoplasms Microgels Plasma samples business.industry Computer science Process Chemistry and Technology Absolute quantification Hand held Loop-mediated isothermal amplification Bioengineering Gene mutation Chip Template Molecular Diagnostic Techniques Carcinoma Non-Small-Cell Lung Humans Digital polymerase chain reaction business Nucleic Acid Amplification Techniques Instrumentation Computer hardware |
| Zdroj: | ACS Sensors. 6:3564-3574 |
| ISSN: | 2379-3694 |
| Popis: | Digital polymerase chain reaction (dPCR) has found widespread applications in molecular diagnosis of various diseases owing to its sensitive single-molecule detection capability. However, the existing dPCR platforms rely on the auxiliary procedure to disperse DNA samples, which needs complicated operation, expensive apparatus, and consumables. Besides, the complex and costly dPCR readers also impede the applications of dPCR for point-of-care testing (POCT). Herein, we developed a portable digital loop-mediated isothermal amplification (dLAMP) platform, integrating a microscale hydrogel (microgel) array chip for sample partition, a miniaturized heater for DNA amplification, and a hand-held reader for digital readout. In the platform, the chip with thousands of isolated microgels holds the capability of self-absorption and partition of DNA samples, thus avoiding auxiliary equipment and professional personnel operations. Using the integrated dLAMP platform, λDNA templates have been quantified with a good linear detection range of 2-1000 copies/μL and a detection limit of 1 copy/μL. As a demonstration, the epidermal growth factor receptor L858R gene mutation, a crucial factor for the susceptibility of the tyrosine kinase inhibitor in non-small-cell lung cancer treatment, has been accurately identified by the dLAMP platform with a spiked plasma sample. This work shows that the developed dLAMP platform provides a low-cost, facile, and user-friendly solution for the absolute quantification of DNA, showing great potential for the POCT of nucleic acids. |
| Databáze: | OpenAIRE |
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