Use this URL to cite or link to this record in EThOS: http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.652800
Title: Diagnostic methods for epidemiological studies of tropical theileriosis
Author: Ilhan, T.
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 2000
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Abstract:
The aim of this study was to develop improved diagnostic techniques for tropical theileriosis caused by T. annulata. These were (i) stage specific indirect enzyme-linked immunosorbent assays (ELISA) to distinguish animals vaccinated with high passage cell lines from those naturally infected in the field and (ii) a highly sensitive and specific polymerase chain reaction (PCR) methodology to detect low level infections in carrier animals. In order to obtain an antigen expressed by the macroschizont stage of the parasite to use in ELISA an attempt was made to isolate a T. annulata gene, which was a homologue of the QP rich protein of T. parva, from a T. annulata l ZAP cDNA library. Three T. annulata genes, NCl, NC2 and NC10, were isolated from the cDNA library by screening with QP cDNA probe. Characterisation of these genes was carried out by Southern, northern, western blot and sequence analyses. An immunogenic recombinant protein, NC10-Ssp13, was obtained from the NC10 gene which was expressed by the macroschizont stage of the parasite. Antisera against the NC10-Ssp13 antigen detected polypeptides both in macroschizonts and host nucleus. The results of this study show that, ELISAs using the Tamr-1, NC10-Ssp13 of Tash-2 recombinant antigens did not reliably distinguish animals vaccinated with the high passage cell line from those that were naturally infected. Both Tamr-1 and Tash-2 ELISAs, however, could serve as powerful diagnostic tools in epidemiological studies of theileriosis, as ELISA systems permit processing of larger numbers of samples than IFAT and give objective results. The nested PCR using primers derived from the gene encoding the 30 kDa major merozoite surface antigen (Tams-1) of T. annulata amplified parasite DNA in blood samples from animals exhibiting low piroplasm parasitaemia. The PCR distinguishes T. annulata from T. buffeli in cattle in T. annulata from T. lestoquardi and B. equi in vector ticks. Such sensitive and specific tests would complement the sero-epidemiological studies of theileriosis and help to target the vaccine produced.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.652800  DOI: Not available
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