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Home / Equine Research Bank / Int J Parasitol / The beta-tubulin gene of Babesia and Theileria parasites is ...
International journal for parasitology2000; 30(11); 1181-1185; doi: 10.1016/s0020-7519(00)00105-3

The beta-tubulin gene of Babesia and Theileria parasites is an informative marker for species discrimination.

Abstract: A fragment of the beta-tubulin gene was polymerase chain reaction (PCR) amplified from genomic DNAs of Babesia bovis, Babesia bigemina, Babesia divergens, Babesia major, Babesia caballi, Babesia equi, Babesia microti, Theileria annulata and Theileria sergenti. Single amplification products were obtained for each of these species, but the size of the amplicons varied from 310 to 460 bp. Sequence analysis revealed that this variation is due to the presence of a single intron, which ranged from 20 to 170 bp. The extensive genetic variability at the beta-tubulin locus has been exploited to develop two types of species identification assays. The first assay can be used on samples containing mostly parasite DNA, like those prepared from infected erythrocytes. Following PCR amplification, the species identification is obtained directly from the size of the products (for Babesia species infecting human or horse) or using a simple PCR-restriction fragment length polymorphism (RFLP) protocol (for Babesia species infecting cattle). The second assay can be used on samples prepared from whole blood, that contain both parasite and host DNAs. In this case, due to the strong conservation of the beta-tubulin gene, co-amplification of a gene fragment from the host DNA was observed. A nested PCR assay was developed for the specific amplification of parasite DNA, using a primer designed to span the exon-intron boundary. Direct identification of Babesia species infecting human and horse is again obtained after the electrophoretic separation of the amplification products, while for Babesia and Theileria species infecting cattle, differentiation is based on a nested PCR-RFLP protocol. These methods may be used for the simultaneous identification of horses and cattle carrying multiple parasites by means of a single PCR or using the PCR-RFLP protocol.
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Publication Date: 2000-10-12 PubMed ID: 11027785DOI: 10.1016/s0020-7519(00)00105-3Google Scholar: Lookup
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  • Journal Article

Summary

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This research article investigates the genetic variability of the beta-tubulin gene in different species of Babesia and Theileria parasites, stating that it can be used as an informative marker for species identification. It discusses two specific methods that exploit this genetic variability to identify what species the parasites belong to.

Methods and Results

  • Genomic DNAs of different species of Babesia and Theileria parasites were subjected to Polymerase Chain Reaction (PCR) to amplify a fragment of their beta-tubulin gene. They found that each species produced unique amplification products and the size of these products varied from species to species
  • The difference in size was caused by the presence of a singular intron, whose size ranged from 20 to 170 bp.
  • Two assays based on the species’ amplification product size and variation at the beta-tubulin gene locus were developed for species identification.

First Assay

  • This assay is applied to samples having high healthy levels of parasite DNA, as obtained from infected erythrocytes.
  • The species identification is done directly from the sizes of the PCR products for Babesia species infecting human or horse or a PCR-restriction fragment length polymorphism (RFLP) protocol for Babesia species infecting cattle.

Second Assay

  • This assay is used on samples that contain both host and parasite DNA and are prepared from whole blood.
  • Due to the strong conservation of the beta-tubulin gene, a fragment of the host DNA is also co-amplified. To overcome this, a nested PCR assay designed to target the exon-intron boundary is used, allowing for the specific amplification of the parasite DNA.
  • Like the first assay, direct species identification is obtained after the separation of the PCR products by electrophoresis. This applies to Babesia species affecting humans and horses. For Babesia and Theileria species infecting cattle, species differentiation is based on a nested PCR-RFLP protocol.

Conclusion

  • The developed methods could be instrumental in identifying horses and cattle infected with multiple parasites simultaneously through a single PCR or using the PCR-RFLP protocol.

Cite This Article

APA
Cacciò S, Cammà C, Onuma M, Severini C. (2000). The beta-tubulin gene of Babesia and Theileria parasites is an informative marker for species discrimination. Int J Parasitol, 30(11), 1181-1185. https://doi.org/10.1016/s0020-7519(00)00105-3

Publication

International journal for parasitology
ISSN: 0020-7519
NlmUniqueID: 0314024
Country: England
Language: English
Volume: 30
Issue: 11
Pages: 1181-1185

Researcher Affiliations

Cacciò, S
  • Laboratory of Parasitology, Istituto Superiore di Sanità, Viale Regina Elena, 299, 00161 Rome, Italy. caccio@iss.it
Cammà, C
    Onuma, M
      Severini, C

        MeSH Terms

        • Animals
        • Babesia / chemistry
        • Babesia / genetics
        • Babesia / isolation & purification
        • Babesiosis / blood
        • Babesiosis / diagnosis
        • Base Sequence
        • Biomarkers
        • Cattle
        • DNA Primers / chemistry
        • DNA, Protozoan / chemistry
        • DNA, Protozoan / isolation & purification
        • Deoxyribonucleases, Type II Site-Specific / chemistry
        • Electrophoresis, Agar Gel
        • Genetic Variation / genetics
        • Horses
        • Humans
        • Introns
        • Molecular Sequence Data
        • Polymerase Chain Reaction / methods
        • Polymorphism, Restriction Fragment Length
        • Sensitivity and Specificity
        • Sequence Analysis, DNA
        • Theileria / chemistry
        • Theileria / genetics
        • Theileria / isolation & purification
        • Theileriasis / blood
        • Theileriasis / diagnosis
        • Tubulin / chemistry
        • Tubulin / genetics

        Citations

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