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Updated: Aug 13, 2026
Design to Implementation Study for Development and Patient Validation of Paper-Based Toehold Switch Diagnostics
Published on: June 17, 2022
A point-of-care testing platform for pathogen nucleic acid analysis utilizing paper-based extraction and portable
Jiahao Li1,2, Chuhan Zhang1, Zongdong Ou3
1Sanya Institute of Nanjing Agricultural University, Joint International Research Laboratory of Animal Health and Food Safety of Ministry of Education, Single Molecule Biochemistry & Biomedicine Laboratory (Sinmolab), Nanjing Agricultural University, Nanjing, 210095, PR China.
This study presents a rapid, 45-minute point-of-care diagnostic platform for animal diseases. The system enables on-site pathogen detection, improving early diagnosis and control efforts.
Area of Science:
- Veterinary Diagnostics
- Molecular Biology
- Point-of-Care Testing
Background:
- Rapid and reliable diagnostics are essential for animal disease control, especially when treatments are limited.
- Conventional nucleic acid detection methods like real-time quantitative polymerase chain reaction (qPCR) are sensitive but time-consuming and require centralized labs.
- Current workflows involve lengthy sample processing (1.5–3 hours) and transportation delays, hindering timely outbreak response.
Purpose of the Study:
- To develop and validate a streamlined, on-site diagnostic platform for rapid animal pathogen detection.
- To overcome the limitations of traditional laboratory-based qPCR workflows, including time and logistical constraints.
- To provide a portable solution for early disease diagnosis in resource-limited settings.
Main Methods:
- Integration of a centrifuge-free, glass fiber paper-based nucleic acid extraction card.
- Utilization of room-temperature-stable lyophilized qPCR reagents.
- Development of a portable qPCR instrument for integrated sample-to-answer analysis.
Main Results:
- The platform delivers results within 45 minutes, significantly reducing turnaround time.
- Achieved high analytical sensitivity with limits of detection of 10 copies/μL for porcine epidemic diarrhea virus (PEDV) and 50 copies/μL for Vibrio parahaemolyticus AHPND (VPAHPND).
- Demonstrated 100% diagnostic concordance with standard laboratory methods in clinical field samples.
Conclusions:
- The developed point-of-care system offers a rapid and accurate solution for on-site animal disease diagnostics.
- This technology streamlines pathogen detection, enabling faster response to outbreaks.
- The platform holds significant potential for improving animal health surveillance, particularly in resource-constrained environments.
