3D Printer-Based PCR Reagent Dispenser with Syringe Pump and Three-Way Valve for Rapid Nucleic Acid Diagnostics

Chi-Uk Han; Jongwon Kim; Mun-Ho Ryu

doi:10.46604/ijeti.2025.14847

Authors

Chi-Uk Han Department of Healthcare Engineering, Jeonbuk National University, Jeonju, Republic of Korea
Jongwon Kim Biomedux Co., Ltd., Gyeonggi, Republic of Korea
Mun-Ho Ryu Division of Biomedical Engineering; Research Center of Healthcare & Welfare Instrument for the Aged, Jeonbuk National University, Jeonju, Republic of Korea

DOI:

https://doi.org/10.46604/ijeti.2025.14847

Keywords:

liquid handling, reagent dispenser, nucleic acid diagnosis, 3D printer

Abstract

This study presents a cost-effective automation solution for preparing polymerase chain reaction (PCR) reagent cartridges used in automated nucleic acid analyzers. Manual preparation is labor-intensive and error-prone, often causing inaccurate volumes and reagent mismatches. To address this, a dispensing system based on open-source 3D printer technology is developed. It incorporates a motion platform and a syringe-based pump, and precisely dispenses reagents into cartridge chambers designed for magnetic DNA extraction and real-time PCR. The system is evaluated for manual inefficiency and error. Dispensing accuracy, assessed gravimetrically using 500 μL of distilled water, shows a relative accuracy of 0.30% and a coefficient of variation (CV) of 2.64%, both within ISO 8655 limits. In terms of efficiency, the system fills a single cartridge chamber in 13.57 seconds, much faster than the approximately 3 minutes required for manual reagent injection. These results highlight the system’s potential to improve throughput and precision in cartridge preparation.

References

Y. Fu, X. Zhou, and D. Xing, “Integrated Paper-Based Detection Chip with Nucleic Acid Extraction and Amplification for Automatic and Sensitive Pathogen Detection,” Sensors and Actuators B: Chemical, vol. 261, pp. 288-296, 2018.

B. H. Kil, J. S. Park, C. Y. Park, Y. S. Kim, and J. D. Kim, “System Architecture for IIoT-Based POC Molecular Diagnostic Device,” Engineering Proceedings, vol. 6, no. 1, article no. 60, 2021.

F. de Mello Malta, D. Amgarten, F. C. Val, M. C. Cervato, B. M. C. de Azevedo, M. de Souza Basqueira, et al., “Mass Molecular Testing for COVID19 Using NGS-Based Technology and a Highly Scalable Workflow,” Scientific Reports, vol. 11, article no. 7122, 2021.

E. S. Goudouris, “Laboratory Diagnosis of COVID-19,” Journal de Pediatria, vol. 97, no. 1, pp. 7-12, 2021.

D. T. P. Thao, V. T. T. Huong, and P. T. H. Thuy, “Application of Realtime RT-PCR Assay for the Detection of E Gene of SARS-CoV-2,” Journal of Control Vaccine and Biologicals, vol. 2, article no. 68, 2022. (In Vietnamese)

I. M. Artika, Y. P. Dewi, I. M. Nainggolan, J. E. Siregar, and U. Antonjaya, “Real-Time Polymerase Chain Reaction: Current Techniques, Applications, and Role in COVID-19 Diagnosis,” Genes, vol. 13, no. 12, article no. 2387, 2022.

B. H. Kil, J. S. Park, M. H. Ryu, C. Y. Park, Y. S. Kim, and J. D. Kim, “Cloud-Based Software Architecture for Fully Automated Point-of-Care Molecular Diagnostic Device,” Sensors, vol. 21, no. 21, article no. 6980, 2021.

S. L. Chen and L. W. Huang, “Using Deep Learning Technology to Realize the Automatic Control Program of Robot Arm Based on Hand Gesture Recognition,” International Journal of Engineering and Technology Innovation, vol. 11, no. 4, pp. 241-250, 2021.

N. Ali, R. de Cássia Pontello Rampazzo, A. D. T. Costa, and M. A. Krieger, “Current Nucleic Acid Extraction Methods and Their Implications to Point-of-Care Diagnostics,” BioMed Research International, vol. 2017, no. 1, article no. 9306564, 2017.

C. E. Reed, J. Fournier, N. Vamvoukas, and S. M. Koza, “Automated Preparation of MS-Sensitive Fluorescently Labeled N-Glycans with a Commercial Pipetting Robot,” Journal of Laboratory Automation, vol. 23, no. 6, pp. 550-559, 2018.

D. C. Florian, M. Odziomek, C. L. Ock, H. Chen, and S. A. Guelcher, “Principles of Computer-Controlled Linear Motion Applied to an Open-Source Affordable Liquid Handler for Automated Micropipetting,” Scientific Reports, vol. 10, article no. 13663, 2020.

M. R. Behrens, H. C. Fuller, E. R. Swist, J. Wu, M. M. Islam, Z. Long, et al., “Open-Source, 3D-Printed Peristaltic Pumps for Small Volume Point-of-Care Liquid Handling,” Scientific Reports, vol. 10, article no. 1543, 2020.

C. E. Anderson, T. Huynh, D. J. Gasperino, L. F. Alonzo, J. L. Cantera, S. P. Harston, et al., “Automated Liquid Handling Robot for Rapid Lateral Flow Assay Development,” Analytical and Bioanalytical Chemistry, vol. 414, no. 8, pp. 2607-2618, 2022.

F. Kong, L. Yuan, Y. F. Zheng, and W. Chen, “Automatic Liquid Handling for Life Science: A Critical Review of the Current State of the Art,” Journal of Laboratory Automation, vol. 17, no. 3, pp. 169-185, 2012.

F. Nejatimoharrami, A. Faina, and K. Stoy, “New Capabilities of EvoBot: A Modular, Open-Source Liquid-Handling Robot,” Journal of Laboratory Automation, vol. 22, no. 5, pp. 500-506, 2017.

L. C. Gerber, A. Calasanz-Kaiser, L. Hyman, K. Voitiuk, U. Patil, and I. H. Riedel-Kruse, “Liquid-Handling Lego Robots and Experiments for STEM Education and Research,” PLoS Biology, vol. 15, no. 3, article no. e2001413, 2017.

F. Barthels, U. Barthels, M. Schwickert, and T. Schirmeister, “FINDUS: An Open-Source 3D Printable Liquid-Handling Workstation for Laboratory Automation in Life Sciences,” Journal of Laboratory Automation, vol. 25, no. 2, pp. 190-199, 2019.

A. Kopyl, Y. Yew, J. W. Ong, T. Hiscox, C. Young, M. Muradoglu, et al., “Automated Liquid Handler from a 3D Printer,” Journal of Chemical Education, vol. 101, no. 2, pp. 640-646, 2024.

E. E. A. W. Councill, N. B. Axtell, T. Truong, Y. Liang, A. L. Aposhian, K. G. I. Webber, et al., “Adapting a Low-Cost and Open-Source Commercial Pipetting Robot for Nanoliter Liquid Handling,” Journal of Laboratory Automation, vol. 26, no. 3, pp. 311-319, 2020.

G. Gome, J. Waksberg, A. Grishko, I. Y. Wald, and O. Zuckerman, “OpenLH: Open Liquid-Handling System for Creative Experimentation with Biology,” Proceedings of the Thirteenth International Conference on Tangible, Embedded, and Embodied Interaction, pp. 55-64, 2019.

M. Storch, M. C. Haines, and G. S. Baldwin, “DNA-BOT: A Low-Cost, Automated DNA Assembly Platform for Synthetic Biology,” Synthetic Biology, vol. 5, no. 1, article no. ysaa010, 2020.

H. Tegally, J. E. San, J. Giandhari, and T. de Oliveira, “Unlocking the Efficiency of Genomics Laboratories with Robotic Liquid-Handling,” BMC Genomics, vol. 21, article no. 729, 2020.

B. Kozub, S. Gądek, B. Tyliszczak, L. Wojnar, and K. Korniejenko, “Leveraging 3D Printing Capability for Geopolymer Composites Based on Fly Ash with Cotton Fibers Addition,” International Journal of Engineering and Technology Innovation, vol. 14, no. 3, pp. 231-243, 2024.

Piston-Operated Volumetric Apparatus, British Standards 8655, 2002.

K. U. Krishnan, M. Adisesh, L. Navaneethakrishnan, and R. Manjunathan, “Calibration of Micropipettes through Gravimetric Solution and Its Beneficial Impact on Research,” BioTechnology: An Indian Journal, vol. 15, no. 4, article no. 195, 2019.