ArticleFrontiers in cellular and infection microbiology2025
The updated duplex fluorescence quantitative RT-PCR assay for simultaneous detection of PRRSV-1 and PRRSV-2.
Article in Frontiers in cellular and infection microbiology, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 7 papers.
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Who cites it
7 citing papers in PubMed.
- Development and Application of a Triplex RT-qPCR Assay for Differentiating Major Lineages of Porcine Reproductive and Respiratory Syndrome Virus.Animals : an open access journal from MDPI · 2026Article
- A Quantitative Real-Time PCR Assay for Detection and Quantification of the Ginseng Alternaria Leaf and Stem Blight PathogenJournal of fungi (Basel, Switzerland) · 2026Article
- The updated one-step multiplex RT-qPCR method for PRRSV classical strains, highly pathogenic strains and NADC30-like strains.Frontiers in microbiology · 2026Article
- Development of a novel duplex crystal digital PCR for the detection of PRRSV-1 and PRRSV-2.Frontiers in cellular and infection microbiology · 2026Article
- Article
- A Triplex Propidium Monoazide (PMA) qPCR Assay Enables Rapid Discrimination of Live Porcine Reproductive and Respiratory Syndrome Viruses.Transboundary and emerging diseases · 2025Article
- Development and application of a qPCR assay forFrontiers in cellular and infection microbiology · 2025Article
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Authors and funding
10 authors.
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Abstract
Introduction: Porcine reproductive and respiratory syndrome (PRRS) is one of the most economically devastating infectious diseases in the global swine industry. With the continuous mutation and recombination of PRRSV, existing detection methods frequently result in false negatives, further complicating the prevention and control of PRRS. Methods: The duplex real-time quantitative RT-PCR (RT-qPCR) for the simultaneous detection of PRRSV-1 and PRRSV-2 was developed by designing specific primers and probes based on the ORF6 gene, which is different from conventional nucleic acid detection methods that are typically based on the ORF7 gene. Results: The method showed high specificity for exclusively detecting PRRSV-1 and PRRSV-2, with no cross-reactivity observed against other porcine pathogens. The limit of detection (LOD) was 8.42 copies for PRRSV-1 and 7.84 copies for PRRSV-2. Intra-assay coefficients of variation (CVs) were 0.22-1.07% and inter-assay CVs were 0.52-1.28%. A total of 356 clinical samples were detected using the developed duplex RT-qPCR and compared to the WOAH-recommended RT-qPCR assay and commercial universal PRRSV RT-qPCR detection kit. The assay established in this study demonstrated higher positivity rates, indicating its superior sensitivity. Discussion: An efficient, sensitive, and accurate method for the detection and differentiation of PRRSV-1 and PRRSV-2 was developed and applied to the detection and monitoring of PRRSV.
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