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

E. N. Barker, S. Tasker, M. J. Day, S. M. Warman, K. Woolley, R. Birtles, K. C. Georges, C. D. Ezeokoli, A. Newaj-Fyzul, M. D. Campbell, O. A.E. Sparagano, S. Cleaveland, C. R. Helps

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Abstract

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 106 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.

Original languageEnglish
Pages (from-to)167-170
Number of pages4
JournalVeterinary Microbiology
Volume140
Issue number1-2
Publication statusPublished - 6 Jan 2010
Externally publishedYes

Fingerprint

Mycoplasma haemocanis
Mycoplasma
Real-Time Polymerase Chain Reaction
quantitative polymerase chain reaction
Dogs
Canidae
dogs
assays
Glyceraldehyde-3-Phosphate Dehydrogenases
glyceraldehyde-3-phosphate dehydrogenase
DNA
plasmids
sampling
Plasmids
Animal Hospitals
Tanzania
veterinary clinics
rRNA Genes
cross reaction
anemia

Bibliographical note

Under a Creative Commons license

Keywords

  • Candidatus Mycoplasma haematoparvum"
  • Mycoplasma haemocanis
  • Polymerase chain reaction
  • Prevalence
  • Ticks

ASJC Scopus subject areas

  • Microbiology
  • veterinary(all)

Cite this

Barker, E. N., Tasker, S., Day, M. J., Warman, S. M., Woolley, K., Birtles, R., ... Helps, C. R. (2010). Development and use of real-time PCR to detect and quantify Mycoplasma haemocanis and "Candidatus Mycoplasma haematoparvum" in dogs. Veterinary Microbiology, 140(1-2), 167-170.

Development and use of real-time PCR to detect and quantify Mycoplasma haemocanis and "Candidatus Mycoplasma haematoparvum" in dogs. / Barker, E. N.; Tasker, S.; Day, M. J.; Warman, S. M.; Woolley, K.; Birtles, R.; Georges, K. C.; Ezeokoli, C. D.; Newaj-Fyzul, A.; Campbell, M. D.; Sparagano, O. A.E.; Cleaveland, S.; Helps, C. R.

In: Veterinary Microbiology, Vol. 140, No. 1-2, 06.01.2010, p. 167-170.

Research output: Contribution to journalArticle

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 2010, 'Development and use of real-time PCR to detect and quantify Mycoplasma haemocanis and "Candidatus Mycoplasma haematoparvum" in dogs' Veterinary Microbiology, vol. 140, no. 1-2, pp. 167-170.
Barker, E. N. ; Tasker, S. ; Day, M. J. ; Warman, S. M. ; Woolley, K. ; Birtles, R. ; Georges, K. C. ; Ezeokoli, C. D. ; Newaj-Fyzul, A. ; Campbell, M. D. ; Sparagano, O. A.E. ; Cleaveland, S. ; Helps, C. R. / Development and use of real-time PCR to detect and quantify Mycoplasma haemocanis and "Candidatus Mycoplasma haematoparvum" in dogs. In: Veterinary Microbiology. 2010 ; Vol. 140, No. 1-2. pp. 167-170.
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