2. Academic Validation
  3. All-In-One OsciDrop Digital PCR System for Automated and Highly Multiplexed Molecular Diagnostics

All-In-One OsciDrop Digital PCR System for Automated and Highly Multiplexed Molecular Diagnostics

  • Adv Sci (Weinh). 2024 Mar 22:e2309557. doi: 10.1002/advs.202309557.
Caiming Li 1 2,  Nan Kang 3,  Shun Ye 4,  Weihang Huang 5,  Xia Wang 6,  Cheng Wang 7,  Yuchen Li 1 8,  Yan-Fei Liu 1 9,  Ying Lan 1,  Liang Ma 10,  Yuhang Zhao 10,  Yong Han 10,  Jun Fu 10,  Danhua Shen 3,  Lianhua Dong 6,  Wenbin Du 1 2 11
Affiliations

Affiliations

  • 1 State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China.
  • 2 College of Life Sciences, University of the Chinese Academy of Sciences, Beijing, 101408, China.
  • 3 Department of Pathology, Peking University People's Hospital, Beijing, 100044, China.
  • 4 Department of Bioengineering, University of California Los Angeles, Los Angeles, CA, 90095, USA.
  • 5 Center for Corpus Research, Department of English Language and Linguistics, University of Birmingham, Edgbaston, Birmingham, B152TT, UK.
  • 6 Center for Advanced Measurement Science, National Institute of Metrology, Beijing, 100013, China.
  • 7 Department of Breast Surgery Huangpu Branch, Shanghai Ninth People's Hospital, Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, 200011, China.
  • 8 Biomedical Sciences College & Shandong Medical Biotechnology Centre, Shandong First Medical University & Shandong Academy of Medical Sciences, Jinan, 250000, China.
  • 9 Research Center for Analytical Sciences, Northeastern University, Shenyang, 110819, China.
  • 10 Maccura Biotechnology Co., Ltd, Chengdu, 611730, China.
  • 11 Savaid Medical School, University of the Chinese Academy of Sciences, Beijing, 101408, China.
PMID: 38516754 DOI: 10.1002/advs.202309557
Abstract

Digital PCR (dPCR) holds immense potential for precisely detecting nucleic acid markers essential for personalized medicine. However, its broader application is hindered by high consumable costs, complex procedures, and restricted multiplexing capabilities. To address these challenges, an all-in-one dPCR system is introduced that eliminates the need for microfabricated chips, offering fully automated operations and enhanced multiplexing capabilities. Using this innovative oscillation-induced droplet generation technique, OsciDrop, this system supports a comprehensive dPCR workflow, including precise liquid handling, pipette-based droplet printing, in situ thermocycling, multicolor fluorescence imaging, and machine learning-driven analysis. The system's reliability is demonstrated by quantifying reference Materials and evaluating HER2 copy number variation in breast Cancer. Its multiplexing capability is showcased with a quadruplex dPCR assay that detects key EGFR mutations, including 19Del, L858R, and T790M in lung Cancer. Moreover, the digital stepwise melting analysis (dSMA) technique is introduced, enabling high-multiplex profiling of seven major EGFR variants spanning 35 subtypes. This innovative dPCR system presents a cost-effective and versatile alternative, overcoming existing limitations and paving the way for transformative advances in precision diagnostics.

Keywords

biomedical engineering; chip‐free microfluidics; digital PCR; molecular diagnostics; multiplex nucleic acid testing.

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