Automated Platform for Rapid and Reproducible Sample Preparations in Point-of-Care(POC) Molecular Diagnostics
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Original Article
Automated Platform for Rapid and Reproducible Sample Preparations in Point-of-Care(POC) Molecular Diagnostics Dongkyu Lee1, Ohwon Kwon 1,*, Kang-Ho Lee1, Jongsu Yun1, Changwon kim1, Joonho Seo1, Jeawan Koo2 & Gyu Man Kim2 Received: 19 March, 2019 / Accepted: 31 May, 2019 / Published online: 5 July, 2019 â’¸The Korean BioChip Society and Springer 2019
Abstract Fully automated system for sample preparation in molecular diagnostics was developed. The small cartridge with multiple chambers and a rotating valve was designed for automated processes such as extraction, separation and purifications during the DNA sample preparation. The automated processes such as injection, mixing, capturing and valve on/off motions were controlled by an affordable, motorized control system. We demonstrated the automated DNA extraction of pathogenic bacteria using this system. The deviations in the purity and real-time polymerase chain reaction (PCR) detection of the automated DNA extraction samples were clearly decreased. The device provided rapid, reliable, and reproducible sample preparation in comparison with manual DNA extraction. This automated platform can be utilized in emergent point-of-care testing of infectious disease and foodborne bacteria. Keywords: Molecular diagnostics, Sample preparation, Nucleic acids, Point-of-care testing, Automated platform
Introduction Development of molecular diagnostic devices has been of great interest due to their high accuracy of detection 1 Department of Medical Devices, Korea Institute of Machinery and Materials (KIMM), Daegu, 42994, Republic of Korea 2 School of Mechanical Engineering, Kyunpook National Univiersity, Daegu, 41566, Republic of Korea *Correspondence and requests for materials should be addressed to O. Kwon ( [email protected])
compared to immunoassay1-3. However, solving problems in the complex processes of sample preparation are always significant challenges for their applications in point-of-care (POC) analysis4-7. To prepare the samples for molecular diagnostics, DNA should be extracted and separated from targets for the successful polymerase chain reaction (PCR) amplification detection. In general, the complex processes of capturing DNA with membrane or magnetic particles, rinsing several times for removal of protein waste, and eluting the extracted DNA in the PCR reagents should be conducted. Although total laboratory automation (TLA) system has implemented in large hospitals for automated sample preparation and detection, these sample preparations are still handled through labor-intensive processes with tedious pipetting in the POC test. However, the many steps of manual preparation often cause the unexpected human errors such as large deviations in sample quality, and need the long turn-over time between each step8-10. Although the DNA purification reagent kits and automation instruments such as GeneXpert® (Cepheid, Sunnyvale, CA), EasyMag (bioMerieux, Durham, NC), EZ1 (Qiagen, Valencia, CA), magNA Pure compact (Roche Diagnostics, Indianapolics, IN), No
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