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Discrimination between six species of Theileria using oligonucleotide probes which detect small subunit ribosomal RNA sequences

Published online by Cambridge University Press:  06 April 2009

B. A. Allsopp ,
H. A. Baylis ,
M. T. E. P. Allsoppi ,
T. Cavalier-Smith ,
R. P. Bishop ,
D. M. Carrington ,
B. Sohanpal  and
P. Spooner
B. A. Allsopp*
Affiliation:
University of Cambridge, Department of Biochemistry, Tennis Court Road, Cambridge CB2 1QW, UK
H. A. Baylis*
Affiliation:
University of Cambridge, Department of Biochemistry, Tennis Court Road, Cambridge CB2 1QW, UK
M. T. E. P. Allsoppi*
Affiliation:
Kings College London, Department of Biophysics, 26–29 Drury Lane, London WC2B 5RL, UK
T. Cavalier-Smith*
Affiliation:
Kings College London, Department of Biophysics, 26–29 Drury Lane, London WC2B 5RL, UK
R. P. Bishop
Affiliation:
International Laboratory for Research on Animal Diseases, P.O. Box 30709, Nairobi, Kenya
D. M. Carrington
Affiliation:
University of Cambridge, Department of Biochemistry, Tennis Court Road, Cambridge CB2 1QW, UK
B. Sohanpal
Affiliation:
International Laboratory for Research on Animal Diseases, P.O. Box 30709, Nairobi, Kenya
P. Spooner
Affiliation:
International Laboratory for Research on Animal Diseases, P.O. Box 30709, Nairobi, Kenya
*
*Onderstepoort Veterinary Institute, Private Bag X5, Onderstepoort 0110, South Africa (address for reprint requests and correspondence).
†University of York, Department of Biology, Heslington, York YO1 5DD, UK.
*Onderstepoort Veterinary Institute, Private Bag X5, Onderstepoort 0110, South Africa (address for reprint requests and correspondence).
‡University of British Columbia, Department of Botany, 6270 University Boulevard, Vancouver V6T 1Z4, Canada.
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Summary

The complete small subunit ribosomal RNA (srRNA) gene of Theileria parva was cloned and sequenced. Two primers were designed which permitted the specific amplification of part of the Theileria srRNA gene from Theileria-infected cell line samples which were predominantly (> 95%) bovine DNA. The sequence of the central (variable) region of the srRNA genes of T. annulata, T. taurotragi, T. mutans and two unidentified parasites referred to as Theileria sp. (buffalo) and Theileria sp. (Marula) were obtained. An alignment of the sequences was generated from which 6 oligonucleotide probes, corresponding to species-specific regions, were designed. These probes were demonstrated to provide unequivocal identification of each of the 6 species either by direct detection of parasite srRNA or by hybridization to amplified parasite srRNA genes. The probes were not able to distinguish buffalo-derived T. parva, the causal agent of Corridor disease, from cattle-derived T. parva, the causal agent of East Coast fever.

Keywords

small ribosomal RNA genesgene amplification (PCR)sequence homologyoligonucleotide probesSporozoa genetics.
Type
Research Article
Information
Parasitology , Volume 107 , Issue 2 , August 1993 , pp. 157 - 165
Copyright
Copyright © Cambridge University Press 1993

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