This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Birkenheuer, A. J.
Right arrow Articles by Breitschwerdt, E. B.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Birkenheuer, A. J.
Right arrow Articles by Breitschwerdt, E. B.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, September 2003, p. 4172-4177, Vol. 41, No. 9
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.9.4172-4177.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Development and Evaluation of a Seminested PCR for Detection and Differentiation of Babesia gibsoni (Asian Genotype) and B. canis DNA in Canine Blood Samples

Adam J. Birkenheuer,1 Michael G. Levy,2 and Edward B. Breitschwerdt1*

Departments of Clinical Sciences,1 Food Animal and Health Resource Management, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina 27606-14282

Received 6 November 2002/ Returned for modification 18 March 2003/ Accepted 8 May 2003

Canine babesiosis has recently been recognized as an emerging infectious disease of dogs in North America. We sought to develop a seminested PCR to detect and differentiate Babesia gibsoni (Asian genotype), B. canis subsp. vogeli, B. canis subsp. canis, and B. canis subsp. rossi DNA in canine blood samples. An outer primer pair was designed to amplify an ~340-bp fragment of the 18S rRNA genes from B. gibsoni (Asian genotype), B. canis subsp. vogeli, B. canis subsp. rossi, and B. canis subsp. canis but not mammalian DNA. Forward primers were designed that would specifically amplify a smaller fragment from each organism in a seminested PCR. The practical limit of detection was 50 organisms/ml of mock-infected EDTA anticoagulated whole blood. The primer pair also amplified an ~370-bp fragment of the B. gibsoni (USA/California genotype) 18S rRNA gene from the blood of an experimentally infected dog with a high percentage of parasitemia. Amplicons were not detected when DNA extracted from the blood of a dog that was naturally infected with Theileria annae at a low percentage of parasitemia was amplified. Due to limited sensitivity, this test is not recommended for the routine diagnosis of B. gibsoni (USA/California genotype) or T. annae. The PCR test did not amplify Toxoplasma gondii, Neospora caninum, Leishmania infantum, Cryptosporidium parvum, or canine DNA under any of the conditions tested. The seminested PCR test was able to detect and discriminate B. gibsoni (Asian genotype), B. canis subsp. vogeli, B. canis subsp. canis, and B. canis subsp. rossi DNA in blood samples from infected dogs.

* Corresponding author. Mailing address: North Carolina State University, College of Veterinary Medicine, 4700 Hillsborough St., Raleigh, NC 27606-1428. Phone: (919) 513-6357. Fax: (919) 513-6336. E-mail: Ed_Breitschwerdt{at}ncsu.edu.

Journal of Clinical Microbiology, September 2003, p. 4172-4177, Vol. 41, No. 9
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.9.4172-4177.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Hegarty, B. C., de Paiva Diniz, P. P. V., Bradley, J. M., Lorentzen, L., Breitschwerdt, E. (2009). Clinical Relevance of Annual Screening Using a Commercial Enzyme-Linked Immunosorbent Assay (SNAP 3Dx) for Canine Ehrlichiosis. Journal of the American Animal Hospital Association 45: 118-124 [Abstract] [Full Text]  
  • Oosthuizen, M. C., Zweygarth, E., Collins, N. E., Troskie, M., Penzhorn, B. L. (2008). Identification of a Novel Babesia sp. from a Sable Antelope (Hippotragus niger Harris, 1838). J. Clin. Microbiol. 46: 2247-2251 [Abstract] [Full Text]  
  • Birkenheuer, A. J., Whittington, J., Neel, J., Large, E., Barger, A., Levy, M. G., Breitschwerdt, E. B. (2006). Molecular characterization of a babesia species identified in a north american raccoon.. J Wildl Dis 42: 375-380 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»