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stablishment of PCR Method for the Detection of Theirelia annulata Based on TaSP Gene

ZHENG Jin-feng1,2, LUO Jin2, XIE Jun-ren2, TIAN Mei-yuan2,YUAN Xiao-song2,WANG Fang-fang2, GUO Jin-xia1, LI Kui1，LIU Guang-yuan2   

  1. 450002,China；2. Key Laboratory of Veterinary Parasitology of Gansu Province，Key Laboratory of Grazing Animal Diseases of Ministry of Agriculture，State Key Laboratory of Veterinary Etiological Biology，Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences, Lanzhou 730046,China)
  • Online:2014-01-20 Published:2014-02-27

Abstract: To establish a PCR method of Theirelia annulata （T. annulata）, a pair of primers was designed to specifically amplify a 393 bp high immunization fragment based on T. annulata surface protein （TaSP） gene conserved region，T. annulata, T. sinensis, T. sergenti, T. uilenbergi, T. luwenshuni, T. ovis, T. equi, B. caballi and B. bovis were tested by PCR with the primers. While T. annulata genome templates were amplified that different concentrations diluted in order to determine the sensitivity of the experiment. 150 bovine blood samples were detected using PCR and microscopic. The PCR results of specificity assay showed that only T. annulata genome template amplified specific fragment. The sensitivity result showed that the minimum dose of T. annulata that could be detected by PCR assay was 10-10. 150 blood samples were detected by PCR assay and microscopic examination of Giemsa-stained blood semars,the results showed that PCR assay was sensitive and specific. It was suitable for the detection of T. annulata.

Key words: Theirelia annulata; TaSP gene; PCR method