Development and preliminary application of PCR-DHPLC assays for detection of the Torque Teno virus / 中国人兽共患病学报

ABSTRACT

In order to identify the Torque Teno virus (TT virus),a PCR-DHPLC assay was performed in this study.Primers specific were selected according to the characteristics of TT virus nucleic acid sequence to conduct PCR,and PCR products assayed by DHPLC.We analyzed the sensitivity,specificity,repeatability of PCR-DHPLC and applied it preliminarily on clinical detection.The specific testing was performed with TTV,HBV,HCV and HEV,no cross reaction were found,and the PCR-DHPLC assays we developed had good specification and nice repeatability.Sensitivity analysis showed that the developed PCR-DHPLC assays could detect 1.0× 101 copy/μL.Then we detected 32 serum samples by this method,real-time PCR and normal PCR at same time.The results showed that 17 TTV positives results could be observed by PCR-DHPLC for 32 samples,it is consistent with real-time PCR test results and 15 positive by normal RT-PCR.PCR-DHPLC assays showed nice specification,sensitivity,repeatability,and could be used in epidemiological investigation.