Applications of Ultra-fast Pulse-controlled PCR (upPCR)
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1)School of Biomedical Engineering (Suzhou), Division of Life Science and Medicine, University of Science and Technology of China, Hefei 230026, China;2)Medical Enzyme Engineering Center, CAS Key Lab of Bio-Medical Diagnostics, Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou 215163, China

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This work was supported by grants from Member Project of Youth Promotion Association of Chinese Academy of Sciences (2022327), The National Natural Science Foundation of China (31900911), Jiangsu Provincial Entrepreneurship and Innovation Talent Project (JSJCRC2021548), and Green Bio-manufacturing Project of the Ministry of Science and Technology (2021YFC2104200).

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    Abstract:

    Under the background of precision medicine and personalized medicine, molecular diagnosis is more and more widely applied in pathogen detection, tumor diagnosis, eugenics and fertility, environmental protection, food safety and other fields, and continues to develop in the direction of molecular point-of-care testing (POCT) with advantages of fast and accurate, low-cost, simple operation. Ultra-fast pulse-controlled PCR (upPCR) is an extension and upgrade of real-time quantitative PCR (qPCR) technology, which uses energy pulses to control metal heating elements (mainly gold nanoparticles) in amplification reactions to complete the rapid heating of the local microenvironment of the solution within a few hundred microseconds, and realize the melting and denaturation of template DNA; after stopping heating, the reaction microenvironment can be rapidly cooled by the surrounding solution down to the extension temperature of the polymerase to achieve amplification of template DNA. A single denaturation-amplification cycle of upPCR is only 1.5-5 s, which is much faster than traditional PCR (about 90 s per cycle), and can thus greatly speed up the PCR reaction. On the basis of retaining the advantages of traditional qPCR such as high sensitivity, high specificity and multiplex detection, ultra-fast pulse control PCR technology adds new advantages such as ultra-fast reaction time (less than 15 min) and simple operation, which is very suitable for molecular POCT scenarios such as home detection and community screening. This paper mainly reviews the principle, core raw materials, equipments and applications of upPCR technology in molecular diagnosis, and discusses the advantages and disadvantages of this technology, as well as future technology development and application trends.

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JING Wei, MA Fu-Qiang. Applications of Ultra-fast Pulse-controlled PCR (upPCR)[J]. Progress in Biochemistry and Biophysics,2023,50(11):2649-2662

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History
  • Received:October 30,2022
  • Revised:October 12,2023
  • Accepted:January 09,2023
  • Online: November 22,2023
  • Published: November 20,2023