Porcine TTSuV Infection Survey Of Jiangxi Province And Porcine Transfusion Transmitted Virus Cloning And Sequence Anaysis

This study, through investigation of TTSuV infection rate and TTSuV cloning andsequence analysis in Jiangxi province, aims at comprehending the infection of Jiangxiswine herd in early and recent time, in order to lay the foundation for future epidemiologyof swine TTSuV virus and in-depth study of gene function.1.porcine TTSuV infection rate survey This study use the investigate method whichcan meanwhile allow high-throughput testing TTSuV1and TTSuV2. We use this duplexTaqman real-time fluorescence quantitative PCR method to detect a total number of1168Jiangxi clinical pig serum samples’ infection rate of TTSuV1and TTSuV2from year2001to2004and year2011to2012. Results showed that the prevalence of two gene typeviruses in all samples wereTTSuV1:72.9%, TTSuV2:69.8%, co-infected rate:53.0%.Result of2001-2004was TTSuV1:73.2%, TTSuV2:71.1%, co-infected rate:53.8%.Result of2011-2012was TTSuV1:72.3%, TTSuV2:68.9%, co-infected rate:53.9%.Itshowed that there is a high TTSuV infection rate in Jiangxi swine farms, There is nosignificant difference on the prevalence of gene type1and2overall, and two geneticsubtypes of co-infection rate is high.2.TTSuV Cloning and homology analysis We designed this two subtypes each apair of primers based on TTSuV1gene sequence (accession number: AB076001) andTTSuV2gene sequence (accession number: AY823991). The length of sequence fragmentTTSuV1-D is about1000bp, sequence fragment TTSuV2-S is about920bp. We selectedstrong positive samples after duplex Taqman real-time fluorescence quantitative PCRdetection to extract total nucleic acid as template. Using the ordinary PCR amplification,byTA cloning built into the pMD18-vector,and transformed it into E.Coli DH5α.Afterplasmid double Enzyme digested validation and sequencing,the results showed that wehave been successed in constructing the TTSuV1and TTSuV2gene into the vector. Bynucleotide homologous comparison and analyze of the isolate strains and related sequencesin GenBank, we concluded that: the gene sequence TTSuV1homology with JiangXi strain(accession number: JX173481) is95.5%, the gene sequence TTSuV2homology withSichuan strain (accession number: HQ204188) is71.4%. It showed that, TTSuV2genetype has a high variability.