Detection of bovine respiratory pathogens using real-time PCR and bead-based technologies
Date
2024
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Abstract
The 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.
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Subject
disease
respiratory
bovine
xTAG
Luminex