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bioRxiv · 10.1101/2025.11.06.687058

Development and Evaluation of Novel Quantitative PCR (qPCR) and Loop-Mediated Isothermal Amplification (LAMP) Assays for Bovine Adenovirus Type 7

Kamel, M.·Davidson, J. L.·Dextre, A.·Ault, A.·Pillai, D.·Koziol, J.·Schoonmaker, J. P.·Johnson, T. A.·Verma, M. S.
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Abstract

Herein, we report the development of molecular assays for the specific detection of bovine adenovirus type 7 (BAV-7), a prevalent pathogen associated with bovine respiratory disease (BRD). To overcome the limitations of the current diagnostic methods, we developed and optimized a TaqMan quantitative polymerase chain reaction (qPCR) assay with a limit of detection (LOD) of 50 copies per reaction (10 copies/{micro}L) and 100% analytical specificity, showing no cross-reactivity with nine other viral pathogens commonly linked to BRD. Additionally, we designed a fluorescent quantitative loop-mediated isothermal amplification (qLAMP) assay demonstrating equivalent LOD and specificity. Both assays were evaluated using 24 bovine nasal swabs from BRD animals, revealing a 92% agreement between qPCR and qLAMP results. These assays provide powerful tools for advancing our understanding of BAV-7 epidemiology and improving disease management strategies. The qLAMP assay, with its rapid and isothermal amplification, presents potential for future development as a field-deployable diagnostic tool to enable timely detection of BAV-7 in cattle.

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Kamel, M., Davidson, J. L., Dextre, A., Ault, A., Pillai, D., Koziol, J., Schoonmaker, J. P., Johnson, T. A., Verma, M. S.. 2025-11-07. Development and Evaluation of Novel Quantitative PCR (qPCR) and Loop-Mediated Isothermal Amplification (LAMP) Assays for Bovine Adenovirus Type 7. https://doi.org/10.1101/2025.11.06.687058

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