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Development and use of real-time PCR to detect and quantify Mycoplasma haemocanis and "Candidatus Mycoplasma haematoparvum" in dogs

Lookup NU author(s): Dr Olivier Sparagano


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Two canine haemoplasma species have been recognised to date; Mycoplasma haemocanis (Mhc), which has been associated with anaemia in splenectomised or immunocompromised dogs, and "Candidatus Mycoplasma haematoparvum" (CMhp), recently described in an anaemic splenectomised dog undergoing chemotherapy. The study aim was to develop quantitative real-time PCR assays (qPCRs) incorporating an endogenous internal control to detect Mhc and CMhp and to apply these assays to DNA samples extracted from canine blood collected in Northern Tanzania (n = 100) and from dogs presented to a Trinidadian veterinary hospital (n = 185). QPCRs specific for Mhc and CMhp were designed using 16S rRNA gene sequence data, and each was duplexed with an assay specific for canine glyceraldehyde-3-phosphate dehydrogenase (GAPDH). The assays detected <= 10 copies of a sequence-specific haemoplasma plasmid per reaction and neither assay showed cross-reactivity with 10(6) copies of the sequence-specific plasmid from the non-target canine haemoplasma species. Nineteen of the 100 Tanzanian samples (19%) were positive for Mhc alone and one (1%) was dually infected. One Trinidadian sample was negative for canine GAPDH DNA and was excluded from the study. Of the 184 remaining Trinidadian samples, nine (4.9%) were positive for Mhc alone, five (2.7%) for CMhp alone, and two (1.1%) dually infected. This is the first report of canine haemoplasma qPCR assays that use an internal control to confirm the presence of amplifiable sample DNA, and their application to prevalence studies. Mhc was the most commonly detected canine haemoplasma species. (C) 2009 Elsevier B.V. All rights reserved.

Publication metadata

Author(s): Barker EN, Tasker S, Day MJ, Warman SM, Woolley K, Birtles R, Georges KC, Ezeokoli CD, Newaj-Fyzul A, Campbell MD, Sparagano OAE, Cleaveland S, Helps CR

Publication type: Article

Publication status: Published

Journal: Veterinary Microbiology

Year: 2010

Volume: 140

Issue: 1-2

Pages: 167-170

ISSN (print): 0378-1135

ISSN (electronic): 1873-2542

Publisher: Elsevier


DOI: 10.1016/j.vetmic.2009.07.006


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Funder referenceFunder name
BSAVA PetSavers
Pfizer Health Limited
University of Bristol
Waltham Foundation
23601University of the West Indies
NSF/DEB0225453National Institutes of Health/National Science Foundation
WT077718Wellcome Trust