ArticlePloS one2013
Development and validation of a multiplex, real-time RT PCR assay for the simultaneous detection of classical and African swine fever viruses.
Article in PloS one, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 47 papers, 1 of them a synthesis that pooled it.
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Who cites it
47 citing papers in PubMed, 1 synthesis or guideline pooled it, 82 citations in OpenAlex.
- Practical Guidance for Clinical Microbiology Laboratories: Viruses Causing Acute Respiratory Tract Infections.Clinical microbiology reviews · 2019Guideline
- African swine fever (asf) control and repopulation strategies in tropical condition: effectiveness of cleaning and disinfection protocols.Tropical animal health and production · 2026Article
- Advancements in Modern Nucleic Acid-Based Multiplex Testing Methodologies for the Diagnosis of Swine Infectious Diseases.Veterinary sciences · 2025Review
- Development and application of a triplex real-time PCR method for the detection ofMicrobiology spectrum · 2025Article
- Comparison of DNA extraction methods for detecting African swine fever virus in feed and environmental samples.Frontiers in veterinary science · 2025Article
- A Novel and Rapid Selective Viral Genome Amplification and Sequencing Method for African Swine Fever Virus.Viruses · 2024Article
- Antiviral drugs in animal-derived matrices: A review.Heliyon · 2024Review
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- Article
- Rapid Differential Detection of Wild-Type Classical Swine Fever Virus and Hog Cholera Lapinized Virus Vaccines by TaqMan MGB-Based Dual One-Step Real-Time RT-PCR.Veterinary sciences · 2024Article
- Development of a novel sensitive single-tube nested PCR assay for the detection of African swine fever virus.Archives of virology · 2024Article
- African Swine Fever Diagnosis in Africa: Challenges and Opportunities.Pathogens (Basel, Switzerland) · 2024Review
- Overview of Modern Commercial Kits for Laboratory Diagnosis of African Swine Fever and Swine Influenza A Viruses.Viruses · 2024Review
- A novel multiplex RT-qPCR assay for simultaneous detection of bovine norovirus, torovirus, and kobuvirus in fecal samples from diarrheic calves.Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc · 2023Article
- Article
- Internal Validation of the ASFV MONODOSE dtec-qPCR Kit for African Swine Fever Virus Detection under the UNE-EN ISO/IEC 17025:2005 Criteria.Veterinary sciences · 2023Article
- Development of a TaqMan-Probe-Based Multiplex Real-Time PCR for the Simultaneous Detection of African Swine Fever Virus, Porcine Circovirus 2, and Pseudorabies Virus in East China from 2020 to 2022.Veterinary sciences · 2023Article
- Development of a multiplex qRT-PCR assay for detection of classical swine fever virus, African swine fever virus, andFrontiers in veterinary science · 2023Article
- African Swine Fever Virus Load in Hematophagous Dipterans Collected in Outbreaks from Romania: Risk Factors and Implications.Transboundary and emerging diseases · 2023Article
- Point-of-Care Testing for Sensitive Detection of the African Swine Fever Virus Genome.Viruses · 2022Article
Corrections and comments
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Authors and funding
5 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
A single-step, multiplex, real-time polymerase chain reaction (RT-PCR) was developed for the simultaneous and differential laboratory diagnosis of Classical swine fever virus (CSFV) and African swine fever virus (ASFV) alongside an exogenous internal control RNA (IC-RNA). Combining a single extraction methodology and primer and probe sets for detection of the three target nucleic acids CSFV, ASFV and IC-RNA, had no effect on the analytical sensitivity of the assay and the new triplex RT-PCR was comparable to standard PCR techniques for CSFV and ASFV diagnosis. After optimisation the assay had a detection limit of 5 CSFV genome copies and 22 ASFV genome copies. Analytical specificity of the triplex assay was validated using a panel of viruses representing 9 of the 11 CSFV subgenotypes, at least 8 of the 22 ASFV genotypes as well as non-CSFV pestiviruses. Positive and negative clinical samples from animals infected experimentally, due to field exposure or collected from the UK which is free from both swine diseases, were used to evaluate the diagnostic sensitivity and specificity for detection of both viruses. The diagnostic sensitivity was 100% for both viruses whilst diagnostic specificity estimates were 100% for CSFV detection and 97.3% for ASFV detection. The inclusion of a heterologous internal control allowed identification of false negative results, which occurred at a higher level than expected. The triplex assay described here offers a valuable new tool for the differential detection of the causative viruses of two clinically indistinguishable porcine diseases, whose geographical occurrence is increasingly overlapping.
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