DEVELOPMENT OF DNA PROBES FOR THE DIAGNOSIS OF HEMOPARASITE INFECTIONS.

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In order to establish a method of diagnosis for hemoparasite infections, we have developed DNA probes and polymerase chain reaction (PCR) for bovine theileriosis. Two genes of Theileria sergenti encoding p32, a major piroplasm surface protein, and p70, a family of heat shock protein, were cloned and sequenced. The cDNA clone of p32, as well as cloned genomic DNA fragment, was used as a DNA probe. We also developed PCR for bovine theileriosis. These techniques provide sensitive, specific and early diagnosis methods for bovine theileriosis. Genetic diversity of the parasite was revealed by using these DNA probes. Allele-specific PCR was developed to detect and differentiate three allelic forms of p32. By this method, it was revealed that most of fields isolates consisted of a mixture of two to three parasite populations bearing different p32 allele and that population changes might occur during persistent infection and infection cycle between mammalian hosts and vector ticks. The PCR detection system developed in this study will be commercialized.