期刊
TALANTA
卷 250, 期 -, 页码 -出版社
ELSEVIER
DOI: 10.1016/j.talanta.2022.123725
关键词
Digital PCR; Direct detection; Multiplex detection; EGFR mutations; ctDNA
资金
- Natural Sci- ence Foundation of Zhejiang Province [LQ22F010005]
- National Natural Science Foundation of China [32071481]
- Open Research Project of the State Key Laboratory of Industrial Control Technology, Zhejiang University, China [ICT2022B54]
This study developed a digital PCR chip with multiplex capability and established a direct amplification detection method without nucleic acid extraction. The chip allows for accurate multiplex detection of ctDNA and eliminates the disadvantages of the extraction process.
Digital PCR is a sensitive detection method, which has important applicability in liquid biopsy through the measurement of ctDNA. However, the current sample pre-processing of ctDNA and the multiplex detection capability of digital PCR have limitations. In view of the above two aspects, we developed a digital PCR chip with multiplex capability and established a direct amplification detection method without nucleic acid extraction. Through the design and processing of the chip, we established a self-priming multiplex digital PCR chip, which can detect 4 targets using single fluorescence. This method can be applied to most digital PCR chips. In addition, we used the plasma of lung cancer patients to establish a direct digital PCR detection method based on the chip, thereby avoiding disadvantages caused by the ctDNA extraction process. As a proof of concept, we prepared blood plasma samples with different concentration of ctDNA to prove the chip's multiplex detection capabilities and the results suggested that this multiplex digital PCR is accurate. Overall, our platform provides a novel and promising option for the detection of ctDNA.
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