4.7 Article

Direct digital polymerase chain reaction chip for the detection of EGFR T790M mutation in plasma

Journal

TALANTA
Volume 237, Issue -, Pages -

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ELSEVIER
DOI: 10.1016/j.talanta.2021.122977

Keywords

Digital PCR; Direct detection; Microfluidics chip; EGFR T790M

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A direct digital chip was developed for nucleic acid detection without extraction and purification, showing high accuracy in clinical sample testing. This chip provides fast sampling and accurate quantification, with potential applications in diagnosis, screening, and research.
Nucleic acid extraction and purification before amplification is considered an essential step for nucleic acid amplification testing. However, this may cause losses or introduce errors that can lead to inaccurate results, especially when using samples with a small nucleic acid concentration. Here, we developed a direct digital chip that enabled us to detect nucleic acid without DNA extraction and purification. We have developed a self-priming liquid-dispensing digital PCR chip that does not require any external power. This is a robust anti-evaporation digital PCR chip with fast sampling and accurate quantification performance. Using this chip, we have estab-lished an on-chip direct nucleic acid amplification method that does not require nucleic acid extraction and purification for liquid biopsy samples. In order to verify the feasibility of this chip for clinical samples, we detected the EGFR T790M mutation from plasma. Results showed that EGFR T790M mutation could be detected with an accuracy of 100% and a sensitivity of 0.01%. Without nucleic acid extraction and purification, the assay avoids complex pre-processing, thus saving time and achieving precise quantification. We expect our direct digital PCR chip to have practical applications in diagnosis, screening, and research, especially in resource-deprived regions.

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