A sensitive and accurate recombinase polymerase amplification assay for detection of the primary bacterial pathogens causing bovine respiratory disease

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creativework.keywords - en

recombinase polymerase amplification

bovine respiratory disease

antimicrobial resistance

integrative conjugative element

creativework.keywords - fr

amplification de la recombinase polymérase

complexe respiratoire bovine

résistance antimicrobienne

élément conjugué intégratif

dc.contributor.author

Conrad, Cheyenne C.

Daher, Rana K.

Stanford, Kim

Amoako, Kingsley K.

Boissinot, Maurice

Bergeron, Michel G.

Alexander, Trevor

Cook, Shaun

Ralston, Brenda

Zaheer, Rahat

Niu, Yan D.

McAllister, Tim

dc.date.accessioned

2023-04-17T13:32:06Z

dc.date.available

2023-04-17T13:32:06Z

dc.date.issued

2020-04-22

dc.description.abstract - en

Rapid and accurate diagnosis of bovine respiratory disease (BRD) presents a substantial challenge to the North American cattle industry. Here we utilize recombinase polymerase amplification (RPA), a fast and sensitive isothermal DNA-based technology for the detection of four BRD pathogens (Mannheimia haemolytica, Pasteurella multocida, Histophilus somni, Mycoplasma bovis), genes coding antimicrobial resistance (AMR) and integrative conjugative elements (ICE) which can harbor AMR genes. Eleven RPA assays were designed and validated including: a) one conventional species-specific multiplex assay targeting the 4 BRD pathogens, b) two species-specific real-time multiplex RPA assays targeting M. haemolytica/M. bovis and P. multocida/H. somni, respectively with a novel competitive internal amplification control, c) seven conventional assays targeting AMR genes (tetH, tetR, msrE, mphE, sul2, floR, erm42), and d) one real-time assay targeting ICE. Each real-time RPA assay was tested on 100 deep nasopharyngeal swabs (DNPS) collected from feedlot cattle previously assessed for targets using either culture methods and/or polymerase chain reaction (PCR) verification (TC-PCR). The developed RPA assays enabled sensitive and accurate identification of BRD agents and AMR/ICE genes directly from DNPS, in a shorter period than TC-PCR, showing considerable promise as a tool for point-of-care identification of BRD pathogens and antimicrobial resistance genes.

dc.identifier.citation

Conrad, C. C., Daher, R. K., Stanford, K., Amoako, K. K., Boissinot, M., Bergeron, M. G., Alexander, T., Cook, S., Ralston, B., Zaheer, R., Niu, Y. D., & McAllister, T. (2020). A sensitive and accurate recombinase polymerase amplification assay for detection of the primary bacterial pathogens causing bovine respiratory disease. Frontiers in Veterinary Science, 7. https://doi.org/10.3389/fvets.2020.00208

dc.identifier.doi

https://doi.org/10.3389/fvets.2020.00208

dc.identifier.issn

2297-1769

dc.identifier.uri

https://open-science.canada.ca/handle/123456789/186

dc.language.iso

en

dc.publisher

Frontiers Media

dc.rights.openaccesslevel - en

Gold

dc.rights.openaccesslevel - fr

Or

dc.subject - en

Agriculture

dc.subject - fr

Agriculture

dc.subject.en - en

Agriculture

dc.subject.fr - fr

Agriculture

dc.title - en

A sensitive and accurate recombinase polymerase amplification assay for detection of the primary bacterial pathogens causing bovine respiratory disease

dc.title.fosrctranslation - fr

A sensitive and accurate recombinase polymerase amplification assay for detection of the primary bacterial pathogens causing bovine respiratory disease

dc.type - en

Article

dc.type - fr

Article

local.article.journaltitle

Frontiers in Veterinary Science

local.article.journalvolume

7

local.peerreview - en

Yes

local.peerreview - fr

Oui

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