Abstract:

To develop a single-tube nested PCR assay for detection of goat-derived Theileria annulata. According to the published Theileria annulata Tams1 gene sequence in Genbank, two pairs of primers were designed to develop nested PCR assay for detecting the disease caused by goat Theileria annulata. The results of verification experiments were shown that the targeted gene fragment was 476 bp in length and homology of nested PCR product was 93%~98%, and this detection method had no cross reaction with Toxoplasma, Neospora caninum, Eimeria, Eperythrozoon suis, and the sensitivity of the assay reached to 12 fg of DNA. The 40.4% of positive rate in 127 clinical sample were detected by the single-tube nested PCR assay, higher than those by routine PCR and microscopic examination. The results revealed that the single-tube nested PCR assay had higher specificity, sensitivity and it could be an effective detection method of goat Theileria annulata.