Journal article
PCR methods for the discrimination of Babesia bovis isolates
Veterinary Parasitology, Vol.71(4), pp.223-237
1997
Abstract
Three different polymerase chain reaction assays for the typing of isolates of Babesia bovis have been developed and compared with a hybridisation based method. Primers were designed within conserved regions flanking the variable length tandem repeats of the Bv80 and BvVA1 genes. For the long array of repeats in BvVA1, up to 7.5 kb, a modified long template PCR method was developed. The assays were compared using ten independent isolates of Babesia bovis. Using the BvVA1 and Bv80 PCR assays, 13 and 10 genotypes could be discriminated, respectively, with some isolates containing several genotypes. Combining the two PCR assays, 17 genotypes were identified within the ten Babesia bovis isolates. Whilst simpler and requiring less DNA, the BvVA1 PCR analysis exhibited significant bias towards some genotypes of the BvVA1 repeats. Further discrimination of BvVA1 PCR products was achieved using AccI digests producing population specific ladders. Genomic DNA fingerprints were also generated by PCR of DNA using an arbitrary primer (randomly amplified polymorphic DNA, RAPD) revealing polymorphic genotypes that were isolate specific. No amplification of host DNA resulted from any of the three PCR procedures. Babesia bigemina DNA was not amplified by the Bv80 or BvVA1 primers. Applications demonstrating changes in composition of populations of Babesia bovis parasites during attenuation and prolonged culture maintanance are described.
Details
- Title
- PCR methods for the discrimination of Babesia bovis isolates
- Authors/Creators
- A.E. Lew (Author/Creator) - Animal Research InstituteB.P. Dalrymple (Author/Creator) - Commonwealth Scientific and Industrial Research OrganisationP.J. Jeston (Author/Creator) - Animal Research InstituteR.E. Bock (Author/Creator) - Department of Environment, Land, Water and Planning
- Publication Details
- Veterinary Parasitology, Vol.71(4), pp.223-237
- Publisher
- Elsevier BV
- Identifiers
- 991005542296407891
- Copyright
- © 1997 Published by Elsevier B.V.
- Murdoch Affiliation
- Murdoch University
- Language
- English
- Resource Type
- Journal article
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- Collaboration types
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- Citation topics
- 1 Clinical & Life Sciences
- 1.258 Zoonotic Diseases
- 1.258.227 Tick-borne Pathogens
- Web Of Science research areas
- Parasitology
- Veterinary Sciences
- ESI research areas
- Plant & Animal Science