ArticleJournal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc2018
Development of a multiplex real-time PCR assay using two thermocycling platforms for detection of major bacterial pathogens associated with bovine respiratory disease complex from clinical samples.
Article in Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 30 papers.
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Who cites it
30 citing papers in PubMed, 47 citations in OpenAlex.
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- Molecular investigation and viral load analysis of bovine respiratory syncytial virus in cattle with bovine respiratory disease complex in Inner Mongolia, China.Frontiers in cellular and infection microbiology · 2026Article
- Novel Recombinase Polymerase Amplification Assay Is Sensitive for Detection of Macrolide Resistance Genes Relevant to Bovine Respiratory Disease Management in Feedlot Calves.Veterinary sciences · 2025Article
- Cross-Species Transmission at the Wildlife-Livestock Interface: A Case Study of Epidemiological Inference From Mule Deer GPS Collar Data.Ecology and evolution · 2025Article
- Survey ofCanadian journal of veterinary research = Revue canadienne de recherche veterinaire · 2025Article
- Aggregate Sampling to Detect Pathogens and Antimicrobial Resistance Genes Associated with Bovine Respiratory Disease in US Feedlots: A Pilot Study.Veterinary sciences · 2025Article
- Exploring the diagnostic landscape ofFrontiers in microbiology · 2025Review
- Use of Subtherapeutic Tylvalosin AgainstTransboundary and emerging diseases · 2025Article
- Comparing Occurrence of Bovine Respiratory Pathogens Detected by High-Throughput Real-Time PCR in Nasal Swabs and Non-Endoscopic Bronchoalveolar Lavage Samples from Dairy and Veal Calves.Pathogens (Basel, Switzerland) · 2024Article
- Agreement among deep nasopharyngeal sampling culture results for 3 different swab types in preweaning dairy calves.JDS communications · 2024Article
- Establishment of a duplex TaqMan-based real time RT-PCR assay for simultaneous detection of BRSV and BVDV.Frontiers in veterinary science · 2024Article
- Article
- Transcript Characteristics on the Susceptibility Difference of Bovine Respiratory Disease.International journal of genomics · 2023Article
- Efficiency-corrected PCR quantification for identification of prevalence and load of respiratory disease-causing agents in feedlot cattle.Australian veterinary journal · 2022Article
- Potential Facilitation Between a Commensal and a Pathogenic Microbe in a Wildlife Disease.EcoHealth · 2022Article
- Does swab type matter? Comparing methods for Mannheimia haemolytica recovery and upper respiratory microbiome characterization in feedlot cattle.Animal microbiome · 2022Article
- Meta-Analysis of qPCR for Bovine Respiratory Disease Based on MIQE Guidelines.Frontiers in molecular biosciences · 2022Article
- Performance Evaluation of a Novel Ultrafast Molecular Diagnostic Device Integrated With Microfluidic Chips and Dual Temperature Modules.Frontiers in bioengineering and biotechnology · 2022Article
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Authors and funding
6 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Bovine respiratory disease complex (BRDC) is one of the most significant diseases of cattle. Bacterial pathogens involved in BRDC include Mannheimia haemolytica, Mycoplasma bovis, Histophilus somni, and Pasteurella multocida. We developed and evaluated a multiplexed real-time hydrolysis probe (rtPCR) assay using block-based Peltier and rotary-based thermocycling on lung tissue, nasal swabs, and deep nasopharyngeal swabs. The rtPCR results were compared to culture or a gel-based M. bovis PCR using statistical analysis to determine optimum quantification cycle (Cq) cutoffs to maximize agreement. The limits of detection were 1.2-12 CFU/reaction for each pathogen. M. haemolytica was the most prevalent organism detected by rtPCR, and was most frequently found with P. multocida. The rtPCR assay enabled enhanced levels of detection over culture for all pathogens on both thermocycling platforms. The rotary-based thermocycler had significantly lower Cq cutoffs (35.2 vs. 39.7), which maximized agreement with gold standard culture or gel-based PCR results following receiver operating characteristic analysis to maximize sensitivity (Se) and specificity (Sp). However, overall assay Se and Sp were similar on both platforms (80.5% Se, 88.8% Sp vs. 80.1% Se, 88.3% Sp). Implementation of these tests could enhance the detection of these pathogens, and with high-throughput workflows could reduce assay time and provide more rapid results. The assays may be especially valuable in identifying coinfections, given that many more antemortem samples tested in our study were positive for 2 or more pathogens by rtPCR ( n = 125) than were detected using culture alone ( n = 25).
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