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A rapid and sensitive intracellular flow cytometric assay to identify Theileria parva infection within target cells

Published online by Cambridge University Press:  25 September 2007

M. S. ROCCHI ,
K. T. BALLINGALL ,
D. NGUGI ,
N. D. MacHUGH  and
D. J. McKEEVER
M. S. ROCCHI*
Affiliation:
Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik EH26 0PZScotland
K. T. BALLINGALL
Affiliation:
Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik EH26 0PZScotland
D. NGUGI
Affiliation:
Division of Animal Health and Welfare, College of Medicine and Veterinary Medicine, University of Edinburgh, Easter Bush Veterinary Centre, Roslin EH25 9RG, Scotland
N. D. MacHUGH
Affiliation:
Division of Animal Health and Welfare, College of Medicine and Veterinary Medicine, University of Edinburgh, Easter Bush Veterinary Centre, Roslin EH25 9RG, Scotland
D. J. McKEEVER
Affiliation:
Royal Veterinary College, Hawkshead Lane, North Mymms, Hatfield, Hertfordshire AL9 7TA
*
*Corresponding author: Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik EH26 0PZ, Scotland, UK. Tel: 0044 (0)131 4455111. Fax: 0044 (0)131 4456111. E-mail: Mara.Rocchi@Moredun.ac.uk
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Summary

Theileria parva is an intracellular protozoan parasite transmitted by ticks that causes a fatal lymphoproliferative disease of cattle known as East Coast Fever. Vaccination against the disease currently relies on inoculation of the infective sporozoite stage of the parasite and simultaneous treatment with long-acting formulations of oxytetracycline. Sporozoites are maintained as frozen stabilates of triturated infected ticks and the method requires accurate titration of stabilates to determine appropriate dose rates. Titration has traditionally been undertaken in cattle and requires large numbers of animals because of individual variation in susceptibility to infection. An alternative tissue culture-based method is laborious and time consuming. We have developed a flow cytometric method for quantifying the infectivity of sporozoite stabilates in vitro based on the detection of intracellular parasite antigen. The method allows clear identification of parasitized cells with a high degree of sensitivity and specificity. Analysis of infected cells between 48 and 72 h post-infection clearly defines the potential transforming capability of different stabilates.

Keywords

Theileria parvaECFintracellularFACSmonoclonal antibodiesPIMschizont
Type
Research Article
Information
Parasitology , Volume 135 , Issue 2 , February 2008 , pp. 195 - 201
Copyright
Copyright © Cambridge University Press 2007

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References

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