ArticleFrontiers in veterinary science2023
Establishment and application of multiplex droplet digital polymerase chain reaction assay for bovine enterovirus, bovine coronavirus, and bovine rotavirus.
Article in Frontiers in veterinary science, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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11 citing papers in PubMed.
- A DPO-Enhanced Gold Nanoparticle-Assisted PCR Assay for Simultaneous Detection of Bovine Viral Diarrhea Virus, Astrovirus, and Rotavirus.Animals : an open access journal from MDPI · 2026Article
- Developing a recombinase-aided amplification method combined with a lateral flow dipstick assay for rapid triplex detection of bovine coronavirus, infectious bovine rhinotracheitis virus, and bovine viral diarrhea virus.Microbiology spectrum · 2026Article
- Development and Evaluation of Quadruplex Droplet Digital PCR Method to Multiplex Detection of Different Respiratory Pathogens of Chickens.Animals : an open access journal from MDPI · 2026Article
- Detection and molecular characterization of bovine enterovirus E2 from dairy calves with respiratory disease in Urumqi, Xinjiang, China.Frontiers in cellular and infection microbiology · 2026Article
- Diagnostic Methods for Bovine Coronavirus: A Review of Recent Advancements and Challenges.Viruses · 2025Review
- Advances in diagnosis of diseases causing diarrhea in newborn calves.Veterinary research communications · 2025Review
- MVPCR: Multiplex Visual Detection Strategy Based on Ultrafast PCR for Point-of-Care Pathogens Detection Within 10 Min.Phenomics (Cham, Switzerland) · 2025Article
- Advances in Laboratory Diagnosis of Coronavirus Infections in Cattle.Pathogens (Basel, Switzerland) · 2024Review
- Development and evaluation of a triplex droplet digital PCR method for differentiation ofFrontiers in microbiology · 2024Article
- Specific and sensitive detection of bovine coronavirus using CRISPR-Cas13a combined with RT-RAA technology.Frontiers in veterinary science · 2024Article
- A multiplex real-time fluorescence-based quantitative PCR assay for calf diarrhea viruses.Frontiers in microbiology · 2023Article
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10 authors.
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Abstract
Bovine enterovirus (BEV), bovine coronavirus (BCoV), and bovine rotavirus (BRV) are still the major worldwide concerns in the health care of cattle, causing serious economic losses in the livestock industry. It is urgent to establish specific and sensitive methods to detect viruses for the early control of diseases. Droplet digital PCR (ddPCR) has been proposed to effectively detect viral particles, and it does not involve Ct values or standard curves. In this study, we designed specific primers and probes, based on conserved regions of viral genomes, to optimize protocols for a dual ddPCR assay for detecting BCoV and BRV and a multiplex ddPCR assay for BEV, BCoV, and BRV. Sensitivity assays revealed that the lower limit of detection for qPCR was 1,000 copies/μL and for ddPCR for BEV, BCoV, and BRV, 2.7 copies/μL, 1 copy/μL and 2.4 copies/μL, respectively. Studying 82 samples collected from diarrheal calves on a farm, our dual ddPCR method detected BCoV, BRV, and co-infection at rates of 18.29%, 14.63%, and 6.1%, respectively. In contrast, conventional qPCR methods detected BCoV, BRV, and co-infection at rates of 10.98%, 12.2%, and 3.66%, respectively. On the other hand, studying 68 samples from another farm, qPCR detected BCoV, BRV, BEV, and co-infection of BCoV and BEV at rates of 14.49%, 1.45%, 5.80%, and 1.45%, respectively. Our multiplex ddPCR method detected BCoV, BRV, BEV, co-infection of BCoV and BEV, and co-infection of BRV and BEV. at rates of 14.49%, 2.9%, 8.7%, 2.9%, and 1.45%, respectively. Studying 93 samples from another farm, qPCR detected BCoV, BRV, BEV, and co-infection of BCoV and BEV was detected at rates of 5.38%, 1.08%, 18.28%, and 1.08%, respectively. Co-infection of BCoV, BRV, BEV, BCoV, and BEV, and co-infection of BRV and BEV, were detected by multiplex ddPCR methods at rates of 5.38%, 2.15%, 20.45%, 1.08%, and 1.08%, respectively. These results indicated that our optimized dual and multiplex ddPCR methods were more effective than conventional qPCR assays to detect these viral infections.
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