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Detection of bovine respiratory pathogens using real-time PCR and bead-based technologies

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dc.contributor.authorHolmes, Joey, author
dc.contributor.authorPabilonia, Kristy, advisor
dc.contributor.authorMayo, Christie, advisor
dc.contributor.authorEhrhart, Nicole, committee member
dc.date.accessioned2024-09-09T20:51:11Z
dc.date.available2024-09-09T20:51:11Z
dc.date.issued2024
dc.description.abstractThe global cattle industry suffers financial losses of $900 million USD annually from infections caused by respiratory pathogens in the bovine respiratory disease complex (BRD). Accurate and timely detection of BRD pathogens provides cattle producers with a diagnosis so they can institute patient care and prevent pathogen spread. We sought to implement Luminex xTAG technology to detect four pathogens that cause BRD - bovine respiratory syncytial virus (BRSV), bovine viral diarrhea virus (BVDV), bovine herpes virus-1 (BHV-1), and Mycoplasma bovis (M. bovis). We compared singleplex real-time polymerase chain reaction (real-time PCR) to a newly developed xTAG testing protocol. Nucleic acids were extracted from 28 bovine lung samples that previously tested positive on PCR for each of the viral pathogens: BRSV (5), BVDV (5), BHV-1 (5), and M. bovis (5). All samples for BRSV and BHV-1 were detected on xTAG with a mean fluorescent index (MFI) well above 10,000 while detection of BVDV is limited to an MFI of 10,000 and M. bovis is detected inconsistently by xTAG. Lungs from six co-infected animals that tested positive for two BRD pathogens were tested on xTAG and real-time PCR side-by-side, revealing similar findings to the single positive lungs where BHV-1 and BRSV targets are more detectable than BVDV and M. bovis. Spiked pools of all pathogens resulted in MFI decreases as the number of pathogens per sample increases. With proper optimization, Luminex xTAG may be utilized in the veterinary diagnostic setting to circumvent issues with multiplex real-time PCR while maintaining high standards of diagnostic testing.
dc.format.mediumborn digital
dc.format.mediummasters theses
dc.identifierHolmes_colostate_0053N_18498.pdf
dc.identifier.urihttps://hdl.handle.net/10217/239152
dc.languageEnglish
dc.language.isoeng
dc.publisherColorado State University. Libraries
dc.relation.ispartof2020-
dc.rightsCopyright and other restrictions may apply. User is responsible for compliance with all applicable laws. For information about copyright law, please see https://libguides.colostate.edu/copyright.
dc.subjectdisease
dc.subjectrespiratory
dc.subjectbovine
dc.subjectxTAG
dc.subjectLuminex
dc.titleDetection of bovine respiratory pathogens using real-time PCR and bead-based technologies
dc.typeText
dcterms.rights.dplaThis Item is protected by copyright and/or related rights (https://rightsstatements.org/vocab/InC/1.0/). You are free to use this Item in any way that is permitted by the copyright and related rights legislation that applies to your use. For other uses you need to obtain permission from the rights-holder(s).
thesis.degree.disciplineCell and Molecular Biology
thesis.degree.grantorColorado State University
thesis.degree.levelMasters
thesis.degree.nameMaster of Science (M.S.)

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