Moving toward rapid and low-cost point-of-care molecular diagnostics with a repurposed 3D printer and RPA

Authors

Kamfai Chan, Pui-Yan Wong, Chaitanya Parikh, SeasonWong

Keywords

3D printers, Low-resource settings, Nucleic acid extraction, Recombinase polymerase amplification, Foodborne pathogens, Zika virus

Outline

  • Introduction
    • A 3D printer repurposed to perform nucleic acid extraction
    • Need for simple nucleic acid amplification and detection
    • Using a 3D printer for isothermal amplification reactions
    • Using thermoses for RPA reactions
  • Materials and methods
    • Repurposing a 3D printer into an automated NA extraction device
      • Automate extraction and amplification steps using G-code
    • Samples used in this work
      • Bacterial culture conditions
      • Zika virus and saliva specimens
    • Real-time RPA and RT-RPA assays
    • Monitoring and recording the real-time RPA reactions
      • Real-time RPA assay using a benchtop instrument
      • Real-time RPA assay using the 3D printer and a USB-powered portable detector
      • Low-cost real-time RT-RPA assay using the thermos thermal cycler
      • Fluorescent signal detection using smartphone camera
  • Results and discussion
    • 3D printers are fully capable of carrying out high-quality extraction
    • Real-time RPA performed on the 3D printer and ESElog
      • 3D printer movement and scanning
    • Temperature stability of TTC for RPA runs
    • Comparing RPA results from the TTC and a commercial device for RPA
    • Real-time TTC for the real-time detection of L. monocytogenes in milk
  • Conclusions
  • Author contributions
  • Acknowledgements
  • References

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