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Real-time capillary convective PCR based on horizontal thermal convection

  • Xianbo QiuEmail author
  • Jung Il Shu
  • Oktay Baysal
  • Jie Wu
  • Shizhi Qian
  • Shengxiang Ge
  • Ke Li
  • Xiangzhong Ye
  • Ningshao Xia
  • Duli Yu
Research Paper
  • 101 Downloads
Part of the following topical collections:
  1. 2018 International Conference of Microfluidics, Nanofluidics and Lab-on-a-Chip, Beijing, China

Abstract

A real-time, capillary, convective polymerase chain reaction (PCR) system based on horizontal convection is developed and analyzed. The capillary tube reactor is heated at one end in a pseudo-isothermal manner to achieve efficient thermal cycling based on horizontal thermal convection. Mathematical modeling and the in silico simulations indicate that, once consistent temperature gradient along the horizontal capillary tube has been established, a repeatable and continuous circulatory flow in the horizontal directions is created. The formed convection is able to transport PCR reagents through different temperature zones inside the horizontal capillary tube for different reaction stages, that is, DNA denaturing, annealing, and extension in a typical PCR cycle. Furthermore, the effectiveness and efficiency of the horizontal convection for PCR thermal cycling in a capillary tube is confirmed by experimentation. To evaluate the concept of horizontal convective PCR, a compact system, which is able to heat the capillary tube from one end and monitor the fluorescence in situ with a smartphone camera, is developed for real-time amplification. With horizontal thermal convection, influenza A (H1N1) virus nucleic acid targets with a limit of detection (LOD) of 1.0 TCID50/mL can be successfully amplified and detected in 30 min, which is promising for efficient nucleic acid analysis in point-of-care testing.

Keywords

Polymerase chain reaction (PCR) Capillary convective PCR Horizontal thermal convection Real time Point-of-care (POC) testing 

Notes

Funding

The present work was supported by the National Natural Science Foundation of China (No. 81871505, 81371711), National Science and Technology Major Project (2018ZX10732101-001-009), the Fundamental Research Funds for the Central Universities (XK1802-4, PYBZ1830), and the research fund to the top scientific and technological innovation team from Beijing University of Chemical Technology (No. buctylkjcx06).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. Bian X, Jing F, Li G, Fan X, Jia C, Zhou H, Jin Q, Zhao J (2015) A microfluidic droplet digital PCR for simultaneous detection of pathogenic Escherichia coli O157 and Listeria monocytogenes. Biosens Bioelectron 74(4):770CrossRefGoogle Scholar
  2. Boetcher S (2014) Natural convection heat transfer from horizontal cylinders. In: Natural convection from circular cylinders. Springer Briefs in Applied Sciences and Technology. Springer, ChamGoogle Scholar
  3. Cengel YA, Boles MA (2014) Thermodynamics: an engineering approach, 8th edn. McGraw-Hill Higher Education, New YorkGoogle Scholar
  4. Chen Z, Qian S, Abrams WR, Malamud D, Bau HH (2004) Thermosiphon-based PCR reactor: experiment and modeling. Anal Chem 76(13):3707CrossRefGoogle Scholar
  5. Churchill S, Chu H (1975) Correlating equations for laminar and turbulent free convection from a vertical plate. Int J Heat Mass Transf 18(9):1323CrossRefGoogle Scholar
  6. Craw P, Balachandran W (2012) Isothermal nucleic acid amplification technologies for point-of-care diagnostics: a critical review. Lab Chip 12(14):2469CrossRefGoogle Scholar
  7. Greenshields CJ (2017) OpenFOAM User Guide. OpenFOAM FoundationGoogle Scholar
  8. Holman J (2010) Heat transfer. McGraw-Hill Higher Education, 10th ed, New YorkGoogle Scholar
  9. Houssin T, Cramer J, Grojsman R, Bellahsene L, Colas G, Moulet H, Minnella W, Pannetier J, Leberre M, Plecis A, Chen Y (2016) Ultrafast, sensitive and large-volume on-chip real-time PCR for the molecular diagnosis of bacterial and viral infections. Lab Chip 16(8):1401CrossRefGoogle Scholar
  10. Krishnan M, Ugaz VM, Burns MA (2002) PCR in a Rayleigh-Benard convection cell. Science 298:793CrossRefGoogle Scholar
  11. Li Z, Zhao Y, Zhang D, Zhuang S, Yamaguchi Y (2016) The development of a portable buoyancy-driven PCR system and its evaluation by capillary electrophoresis. Sens Actuat B Chem 230:779CrossRefGoogle Scholar
  12. Liao SC, Peng J, Mauk MG, Awasthi S, Song J, Friedman H, Bau HH, Liu C (2016) Smart cup: a minimally-instrumented, smartphone-based point-of-care molecular diagnostic device. Sen Actuat B Chem 229:232CrossRefGoogle Scholar
  13. Liu C, Geva E, Mauk MG, Qiu X, Abrams WR, Malamud D, Curtis K, Owen SM, Bau HH (2011) An isothermal amplification reactor with an integrated isolation membrane for point-of-care detection of infectious diseases. Analyst 136(10):2069CrossRefGoogle Scholar
  14. Priye A, Wong S, Bi Y, Carpio M, Chang J, Coen M, Cope D, Harris J, Johnson J, Keller A, Lim R, Lu S, Millard A, Pangelinan A, Patel N, Smith L, Chan K, Ugaz VM (2016) Lab-on-a-drone: toward pinpoint deployment of smartphone-enabled nucleic acid-based diagnostics for mobile health care. Anal Chem 88(9):4651CrossRefGoogle Scholar
  15. Qiu X, Mauk MG, Chen D, Liu C, Bau HH (2010) A large volume, portable, real-time PCR reactor. Lab Chip 10(22):3170CrossRefGoogle Scholar
  16. Qiu X, Ge S, Gao P, Li K, Yang S, Zhang S, Ye X, Xia N, Qian S (2016) A smartphone-based point-of-care diagnosis of H1N1 with microfluidic convection PCR. Microsyst Technol 23(7):1Google Scholar
  17. Qiu X, Ge S, Gao P, Li K, Yang Y, Zhang S, Ye X, Xia N (2017a) A low-cost and fast real-time PCR system based on capillary convection. J Lab Auto 22(1):13CrossRefGoogle Scholar
  18. Qiu X, Zhang S, Xiang F, Wu D, Guo M, Ge S, Li K, Ye X, Xia N, Qian S (2017b) Instrument-free point-of-care molecular diagnosis of H1N1 based on microfluidic convective PCR. Sens Actuat B Chem 243:738CrossRefGoogle Scholar
  19. Qiu X, Zhang S, Mei L, Wu D, Guo Q, Li K, Ge S, Ye X, Xia N, Mauk MG (2017c) Characterization and analysis of real-time capillary convective PCR toward commercialization. Biomicrofluidics 11(2):024103CrossRefGoogle Scholar
  20. Shu J, Baysal O, Qian S, Qiu X, Wang F (2019) Performance of convective polymerase chain reaction by doubling time. Int J Heat Mass Transf 133:1230CrossRefGoogle Scholar
  21. Son JH, Cho B, Hong SG, Sang HL, Hoxha O, Haack AJ, Lee LP (2015) Ultrafast photonic PCR. LIGHT SCI APPL 4(7):e280CrossRefGoogle Scholar
  22. Sposito A, Hoang V, Devoe DL (2016) Rapid real-time PCR and high resolution melt analysis in a self-filling thermoplastic chip. Lab Chip 16(18):3524CrossRefGoogle Scholar
  23. White F (2011) Fluid mechanics. McGraw-Hill Higher Education, 7th ed, New YorkGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Xianbo Qiu
    • 1
    Email author
  • Jung Il Shu
    • 2
  • Oktay Baysal
    • 2
  • Jie Wu
    • 1
  • Shizhi Qian
    • 2
  • Shengxiang Ge
    • 3
  • Ke Li
    • 4
  • Xiangzhong Ye
    • 4
  • Ningshao Xia
    • 3
  • Duli Yu
    • 1
    • 5
  1. 1.Institute of Microfluidic Chip Development in Biomedical Engineering, College of Information Science and TechnologyBeijing University of Chemical TechnologyBeijingChina
  2. 2.Institute of Micro/NanotechnologyOld Dominion UniversityNorfolkUSA
  3. 3.National Institute of Diagnostics and Vaccine Development in Infectious DiseasesXiamen UniversityXiamenChina
  4. 4.Beijing Wantai Biological Pharmacy Enterprise Co. LtdBeijingChina
  5. 5.Beijing Advanced Innovation Center for Soft Matter Science and EngineeringBeijingChina

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