ArticleViruses2021
Rapid Extraction and Detection of African Swine Fever Virus DNA Based on Isothermal Recombinase Polymerase Amplification Assay.
Article in Viruses, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
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Who cites it
22 citing papers in PubMed.
- A multi-country phase 2 study to evaluate the suitcase lab for rapid detection of SARS-CoV-2 in seven Sub-Saharan African countries: Lessons from the field.Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology · 2023Trial
- Recent advances in isothermal amplification techniques for detection of animal diseases.Frontiers in veterinary science · 2026Review
- The Complexities of African Swine Fever Diagnosis and Control in the Developing World: A Narrative Review Focused on Ghana.Animals : an open access journal from MDPI · 2025Review
- Development of Real-Time and Lateral Flow Dipstick Recombinase Polymerase Amplification Assays for the Rapid Field Diagnosis of MGF-505R Gene-Deleted Mutants of African Swine Fever Virus.Veterinary sciences · 2025Article
- Comparison of DNA extraction methods for detecting African swine fever virus in feed and environmental samples.Frontiers in veterinary science · 2025Article
- Strategic nucleic acid detection approaches for diagnosing African swine fever (ASF): navigating disease dynamics.Veterinary research · 2024Review
- A Low-Cost Handheld Centrifugal Microfluidic System for Multiplexed Visual Detection Based on Isothermal Amplification.Sensors (Basel, Switzerland) · 2024Article
- The Development of a Multienzyme Isothermal Rapid Amplification Assay to Visually Detect Duck Hepatitis B Virus.Veterinary sciences · 2024Article
- Rapid visual detection of Giardia duodenalis in faecal samples using an RPA-CRISPR/Cas12a system.Parasitology research · 2024Article
- African Swine Fever Diagnosis in Africa: Challenges and Opportunities.Pathogens (Basel, Switzerland) · 2024Review
- Stability of GenotubeViruses · 2024Article
- Ultra-rapid and sensitive detection of African swine fever virus using multiple cross displacement amplification combined with nanoparticle-based lateral flow biosensor.Frontiers in microbiology · 2024Article
- A duplex fluorescent quantitative PCR assay to distinguish the genotype I, II and I/II recombinant strains of African swine fever virus in China.Frontiers in veterinary science · 2024Article
- Development and Application of an RPA-Based Rapid Point-of-Care Testing (POCT) Method for the Detection of Feline Panleukopenia Virus.Transboundary and emerging diseases · 2024Article
- Article
- Stability of African swine fever virus genome under different environmental conditions.Veterinary world · 2023Article
- Advanced Strategies for Developing Vaccines and Diagnostic Tools for African Swine Fever.Viruses · 2023Review
- Review
- Development of a highly sensitive point-of-care test for African swine fever that combines EZ-Fast DNA extraction with LAMP detection: Evaluation using naturally infected swine whole blood samples from Vietnam.Veterinary medicine and science · 2023Article
- Efficient detection of African Swine Fever Virus using minimal equipment through a LAMP PCR method.Frontiers in cellular and infection microbiology · 2023Article
Corrections and comments
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Authors and funding
7 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
African swine fever virus (ASFV) is the causative agent of a deadly disease in pigs and is spread rapidly across borders. Samples collected from suspected cases must be sent to the reference laboratory for diagnosis using polymerase chain reaction (PCR). In this study, we aimed to develop a simple DNA isolation step and real-time recombinase polymerase amplification (RPA) assay for rapid detection of ASFV. RPA assay based on the p72 encoding B646L gene of ASFV was established. The assays limit of detection and cross-reactivity were investigated. Diagnostic performance was examined using 73 blood and serum samples. Two extraction approaches were tested: silica-column-based extraction method and simple non-purification DNA isolation (lysis buffer and heating, 70 °C for 20 min). All results were compared with well-established real-time PCR. In a field deployment during a disease outbreak event in Uganda, 20 whole blood samples were tested. The assay's analytical sensitivity was 3.5 DNA copies of molecular standard per µL as determined by probit analysis on eight independent assay runs. The ASFV RPA assay only detected ASFV genotypes. Compared to real-time PCR, RPA diagnostic sensitivity and specificity were 100%. Using the heating/lysis buffer extraction procedure, ASFV-RPA revealed better tolerance to inhibitors than real-time PCR (97% and 38% positivity rate, respectively). In Uganda, infected animals were identified before the appearance of fever. The ASFV-RPA assay is shown to be as sensitive and specific as real-time PCR. Moreover, the combination of the simple extraction protocol allows its use at the point of need to improve control measures.
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