Establishment of a PCR Diagnostic Method for Bovine Theileria annulata Based on TashHN Gene
The synthetic primers were designed and synthesized based on the TashHN gene sequence of Theileria annulata published in Genbank,and a PCR detection method based on the TashHN gene was established.The specificity,sensitivity,and stability of this method were tested,and the established method was used to detect 135 bovine blood samples from various counties in the Guyuan City of Ningxia.The results showed that the established PCR detection method based on the TashHN gene had no cross-reaction with T.sinensis,T.orientalis,Babesia bovis,and B.bigemina;the lowest detectable template concentration was 1.64×102 copies/μL;and the DNA of Theileria annulata could still be detected after being stored in a-20℃freezer for eight months.Five positive samples were detected out of 135 bovine blood samples from Guyuan using this method,which were consistent with those obtained using PCR methods described in existing literature,indicating that the method is accurate and reliable.The established PCR detection method based on the TashHN gene of T.annulata in this experiment has high specificity,sensitivity,and stability,and is suitable for early infection detection of T.annulata in clinical settings and for epidemiological surveys.