Immunological analysis of Babesia equi piroplasm proteins and development of polymerase chain reaction based assay for the diagnosis of equine piroplasmosis Babesia equiピロプラズム蛋白質の免疫学的解析とDNA診断の開発

博士論文

A polymerase chain reaction (PCR) based assay was developed to detect DNA of B abesia equi in naturally and experimentally infected horse blood. The PCR was used to amplify the gene of 37 kDa protein from piroplas m DNA. A pair of synthetic oligonucleotide primers designed from the gene encoding p37 of B. equi amplified a single fragment of approximately 935 bp when the genomic DNAs from USA, South African, Russian and Chinese B. equi stocks were used as template DNAs in PCR. These primers also amplified a single fragment of 1,353 bp from genomic DNA template of Theileria sergenti Chitose and Ikeda stocks. However, no detectable amplifications from DNAs of other B abesia or Theileria species took place under the same conditions. PCR was sensitive enough to detect 10 pg B. equi purified DNA used as template DNA in PCR reaction and could also amplify B. equi DNA in horse blood with 0.001% parasitaemia.