马媾疫锥虫PCR检测方法的研究

Study of a PCR Assay for Detection of Trypanosoma equiperdum

根据GenBank中公布的马媾疫锥虫大环kDNA基因序列,设计合成一对特异性引物,通过对PCR扩增条件的优化,建立了检测马媾疫锥虫的PCR方法。用建立的PCR方法检测马媾疫锥虫、马伊氏锥虫和马焦虫,仅马媾疫锥虫扩增出与实验设计相符的395bp的条带,其余病原体检测均为阴性。灵敏性试验表明:建立的PCR方法最低能检出0.2 pg/μL的马媾疫锥虫DNA。研究建立的马媾疫锥虫PCR方法,具有快速、敏感、特异等优点,可用于临床上马媾疫锥虫感染的检测。

Through the comparison of the kDNA gene of T. Equiperdum from different countries and areas in GenBank, a pair of primers were designed, and a PCR assay was established based on the maxicircle kDNA ofT. Equiperdum.It was shown that T. Equiperdum could be amplified into the specific bands of 395bp by this PCR, but no specific bands of the same sizes were amplified from T. evansi and Theileria equi. The PCR could detect as low as 0.2 pg/μL T. equiperdum DNA. The PCR assay was a quick, sensitive, and specific test for detection ofT. equiperdum, and would be useful for control of the parasite in horses.