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Multiple-probe-assisted DNA capture and amplification for high-throughput African swine fever virus detection
Huicong Wang1, Hongru Pian1, Lihua Fan1
1Department of Biochemistry and Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and School of Basic Medicine, Peking Union Medical College, 5 Dongdan Santiao, Beijing, People's Republic of China.
Applied Microbiology and Biotechnology
|December 28, 2022
Summary
A new method called MADCAT allows rapid and sensitive detection of African swine fever virus (ASFV) without DNA extraction. This high-throughput technology significantly reduces sample-to-result time for effective disease control.
Area of Science:
- Veterinary Virology
- Molecular Diagnostics
- Infectious Disease Surveillance
Background:
- African swine fever (ASF) poses a severe global socio-economic threat to pig populations.
- Rapid and accurate diagnosis is crucial for controlling ASF outbreaks.
- Current diagnostic methods can be time-consuming and complex.
Purpose of the Study:
- To develop a novel, sensitive, simple, and high-throughput method for detecting African swine fever virus (ASFV).
- To reduce the time and complexity of ASF diagnosis.
- To enable large-scale surveillance and monitoring of ASF.
Main Methods:
- Multiple-probe-assisted DNA capture and amplification technology (MADCAT) was developed.
- MADCAT utilizes a direct DNA capture approach on a 96-well plate, bypassing nucleic acid extraction.
- The method involves specific DNA capture followed by amplification for ASFV detection.
Main Results:
- MADCAT achieved a sample-to-result time of approximately 90 minutes for 96 samples, significantly faster than conventional qPCR (3-4 hours).
- The limit of detection (LOD) was 0.5 copies/μL blood, demonstrating 5-fold higher sensitivity than extraction-based qPCR.
- The assay showed 100% specificity against common swine pathogens and 100% diagnostic sensitivity in field samples.
Conclusions:
- MADCAT is a highly sensitive, specific, and rapid diagnostic tool for ASFV detection directly from complex matrices like whole blood.
- The high-throughput nature and simplified workflow make MADCAT suitable for large-scale ASF surveillance and monitoring.
- This technology offers a significant advancement in combating the economic impact of African swine fever.
