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Detection of Theileria annulata in cattle and vector ticks by PCR using the Tams1 gene sequences

Published online by Cambridge University Press:  01 March 2000

E. KIRVAR ,
T. ILHAN ,
F. KATZER ,
P. HOOSHMAND-RAD ,
E. ZWEYGARTH ,
C. GERSTENBERG ,
P. PHIPPS  and
C. G. D. BROWN
E. KIRVAR
Affiliation:
Centre for Tropical Veterinary Medicine, University of Edinburgh, Easter Bush, Roslin, Midlothian EH25 9RG, UK
T. ILHAN
Affiliation:
Centre for Tropical Veterinary Medicine, University of Edinburgh, Easter Bush, Roslin, Midlothian EH25 9RG, UK
F. KATZER
Affiliation:
Department of Veterinary Parasitology, University of Glasgow, Bearsden Road, Glasgow G61 1QH, UK
P. HOOSHMAND-RAD
Affiliation:
Department of Protozoology and Production of Protozoal Vaccines, Razi State Vaccine and Serum Research Institute, PO Box 11365–1558, Tehran, Iran
E. ZWEYGARTH
Affiliation:
Protozoology Division, Onderstepoort Veterinary Institute, Private Bag X5, Onderstepoort, 0110, South Africa
C. GERSTENBERG
Affiliation:
University of Cambridge Department of Clinical Veterinary Medicine, TBA Equine Fertility Unit, Mertoun Paddocks, Newmarket, Suffolk CB8 9BH, UK
P. PHIPPS
Affiliation:
Central Veterinary Laboratory, Parasitology Department, New Haw, Addlestone, Surrey KT15 3NB, UK
C. G. D. BROWN
Affiliation:
Centre for Tropical Veterinary Medicine, University of Edinburgh, Easter Bush, Roslin, Midlothian EH25 9RG, UK
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Abstract

A Polymerase Chain Reaction (PCR) and Southern blot hybridization for the detection of Theileria annulata are described. The PCR used primers amplifying a 785 base-pair fragment of the T. annulata gene which encodes the 30 kDa major merozoite surface antigen, Tams1. The sensitivity of the PCR in bovine blood was 1 piroplasm in 1 μl of blood. T. buffeli, T. parva, Babesia bigemina, B. bovis and B. divergens were not detected. The PCR detected down to 1 infected acinus/tick in resting and partially fed adult Hyalomma anatolicum anatolicum ticks and was negative for T. lestoquardi and T. equi, which are transmitted by this tick but are not infective to cattle. The specificity of the PCR was checked using 30 stocks of T. annulata, all of which were detected. Three stocks of T. lestoquardi, 4 of T. equi and 1 each of T. buffeli, T. parva, B. bigemina, B. bovis and B. divergens were used to ascertain there were no cross-reactions. A nested PCR using separate primers for the first reaction and the same primers for the second reaction detected T. annulata to the same sensitivity and specificity in saponin-extracted DNA samples stored for long periods at −20 °C.

Keywords

Theileria annulatacattleHyalomma anatolicum anatolicumPCR
Type
Research Article
Information
Parasitology , Volume 120 , Issue 3 , March 2000 , pp. 245 - 254
Copyright
2000 Cambridge University Press

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