Application of PCR Assay for Detection of Toxoplasma gondii in Breeding Rhesus Macaques

Objective To establish a rapid,sensitive,specific PCR technique for detection of toxoplasma gondii and applying in breeding rhesus macaques.The results from nested-PCR and single-step PCR assay were compared.Methods Two pairs of primers matched with P30 gene were designed for nested PCR and B1 gene primers were amplified in single-step PCR.Toxoplasma gondii DNA samples were serially tenfold diluted to determine the sensitivity of the nested PCR.150 samples of breeding rhesus macaques from the Institute of Medical Biology were assayed by those two PCR methods.Results The nested PCR was a specific method and its sensitivity was 10~(-3) ng/μL.The results of nested PCR were almost consistent with that of single-step PCR assay,but the positive rate(10%) was higher than that(8.67%) of single-step PCR.Conclusions Both the two PCR methods can be used to detect toxoplasma gondii DNA from monkey samples,and the nested PCR technique is more sensitive than single-step PCR.