We developed a PCR assay for the detection of Babesia odocoilei based on the 18S rRNA gene. Multiple specimens of B. odocoilei were examined, and the assay consistently produced a small specific PCR product of 306 bp. The PCR assay was also challenged with DNA from 13 other Babesia species and 2 Theileria species, originating from 10 different host species; however, nonspecific DNA amplification and multiple banding patterns were observed, and the amplicon banding patterns varied between different isolates of the same species. Sensitivity was determined to be 6.4 pg of DNA, and an estimated 0.0001% parasitism. This assay can be utilized for species-specific differential detection of B. odocoilei.

Biology Centre Institute of Parasitology Czech Academy of Sciences České Budějovice Czech Republic

Canadian Wildlife Health Cooperative Western College of Veterinary Medicine University of Saskatchewan Saskatoon Saskatchewan Canada

CEITEC VFU University of Veterinary and Pharmaceutical Sciences Brno Brno Czech Republic

Department of Botany and Zoology Faculty of Science Masaryk University Brno Czech Republic

Department of Veterinary Pathology Western College of Veterinary Medicine University of Saskatchewan Saskatoon Saskatchewan Canada

National Microbiology Laboratory Public Health Agency of Canada Winnipeg Manitoba Canada

Zobrazit více v PubMed

Aktaş M, et al.. Development of a polymerase chain reaction method for diagnosis of Babesia ovis infection in sheep and goats. Vet Parasitol 2005;133:277–281. PubMed

Ano H, et al.. Detection of Babesia species from infected dog blood by polymerase chain reaction. J Vet Med Sci 2001;63:111–113. PubMed

Bartlett SL, et al.. Diagnosis and treatment of Babesia odocoilei in captive reindeer (Rangifer tarandus tarandus) and recognition of three novel host species. J Zoo Wildl Med 2009;40:152–159. PubMed

Chae JS, et al.. Two Theileria cervi SSU rRNA gene sequence types found in isolates from white-tailed deer and elk in North America. J Wildl Dis 1999;35:458–465. PubMed

Holman PJ, et al.. In vitro cultivation of a Babesia isolated from a white-tailed deer (Odocoileus virginianus). J Parasitol 1988;74:111–115. PubMed

Holman PJ, et al.. Culture isolation and partial characterization of a Babesia sp. from a North American elk (Cervus elaphus). J Wildl Dis 1994;30:460–465. PubMed

Holman PJ, et al.. Antigenic, phenotypic and molecular characterization confirms Babesia odocoilei isolated from three cervids. J Wildl Dis 2000;36:518–530. PubMed

Holman PJ, et al.. Molecular detection of Babesia bovis and Babesia bigemina in white-tailed deer (Odocoileus virginianus) from Tom Green County in central Texas. Vet Parasitol 2011;177:298–304. PubMed

Hrazdilová K, et al.. Diversity of Babesia spp. in cervid ungulates based on the 18S rDNA and cytochrome c oxidase subunit I phylogenies. Infect Genet Evol 2020;77:104060. PubMed

Lockerbie BP, et al.. Use of the polymerase chain reaction assay for the detection of Babesia odocoilei 18S ribosomal RNA in formalin-fixed tissues. J Vet Diagn Invest 2014;26:538–541. PubMed

Maniatis T, et al.. Molecular Cloning: A Laboratory Manual. 2nd ed. Cold Spring Harbor, 1988:E3.

Masuda N, et al.. Analysis of chemical modification of RNA from formalin-fixed samples and optimization of molecular biology applications for such samples. Nucleic Acids Res 1999;27:4436–4443. PubMed PMC

Mathieu A, et al.. Babesia odocoilei as a cause of mortality in captive cervids in Canada. Can Vet J 2018;59:52–58. PubMed PMC

Milnes EL, et al.. Molecular detection of Babesia odocoilei in wild, farmed, and zoo cervids in Ontario, Canada. J Wildl Dis 2019;55:335–342. PubMed

Nakajima R, et al.. Babesia microti-group parasites compared phylogenetically by complete sequencing of the CCTη gene in 36 isolates. J Vet Med Sci 2009;71:55–68. PubMed

Pattullo KM, et al.. Babesia odocoilei infection in a Saskatchewan elk (Cervus elaphus canadensis) herd. J Vet Diagn Invest 2013;25:535–540. PubMed

Perry BD, et al.. Babesia odocoilei Emerson and Wright, 1970 in white-tailed deer, Odocoileus virginianus (Zimmermann), in Virginia. J Wildl Dis 1985;21:149–152. PubMed

Purnell RE, et al.. Separation and recombination of Babesia divergens and Ehrlichia phagocytophila from a field case of redwater from Eire. Vet Rec 1976;99:415–417. PubMed

Purnell RE. Babesiosis in various hosts. In: Ristic M, Kreier JP, eds. Babesiosis. Academic Press, 1981:25–64.

Zahler M, et al.. Detection of a new pathogenic Babesia microti-like species in dogs. Vet Parasitol 2000;89:241–248. PubMed