Establishment Of PCR Detection Assay For TTSuV1TTSuV2and Application

According to the differences between the genotype of Torque teno suis virus(TTSuV). Designed two pairs of PCR primers based on the published TTSuVs sequences in the Genbank TTSuV1(JF694116.1) and TTSuV2(JF694118.1) to detect TTSuV1and TTSuV2.Extracted viral DNA from pig blood samples.amplified TTSuV1and TTSuV2UTR gene.the target fragment were cloned and sequenced.and confirmed by sequence analysis. PCR conditions were optimized by repeatability,sensitivity and specificity assay. Specific experiments to prove that the established PCR method specific the amplification TTSuVs gene sequence and the control group PCV1、PCV2、PRRSV、PRV、PPV couldn’t be amplified. The sensitivity analysis proved that the established assay could detected the lowest concentration of nucleic acids was10-7ng/mL.The assay could effectively detect a small quantity of DNA in the sample. Repeatability tests showed that the established method of high accuracy, and exclude false-positive interference.254serum samples which collected from different farms in Sichuan province were detected by the established methods and maked a preliminary investigation.The survey results show that the254serum samples collected TTSuV1detection rate of18.1%(46/254) and TTSuV2detection rate of39.4%(100/254). TTSuV1and TTSuV2co-infection was 10.2%(26/254).The test results indicated that the TTSuVs infection was ubiquity in Sichuan province and TTSuV2infection has higher positive rate.At the same time.it proved that the PCR assay established in this study has a strong sensitivity and specificity.can be used for largescale clinical testing.The PCR assay established in this study can be fast and accurate detection of porcine TTSuVs with high specificity and sensitivity characteristics, compared to commonly used porcine TTSuVs nest-PCR detection method takes less time, more practical value in clinical. At the same time, this study on the parts of Sichuan TTSuV infection surveys, provide data support for the southwest region of China and even the whole country pig TTSuVs epidemiology survey, but also laid the foundation for porcine TTSuV prevention and control and further study.