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- W2394012938 abstract "This paper described a multiplex RT-PCR to rapidly diagnose the co-infection in samples from pig with reproductive failure symptoms.Three sets of primers were designed,respectively,targeting to the conserved sequences in E2 gene of classical swine fever virus(CSFV),a domain encompassing partial ORF6,ORF7 and 3-terminus UTR of porcine reproductive and respiratory syndrome virus(PRRSV) and E gene of Japanese encephalitis virus(JEV).The expected sizes of RT-PCR amplicons were 288 bp for CSFV,430 bp for PRRSV and 1 015 bp for JEV,respectively.RNA was extracted from pure culture of the viruses,followed by the synthesis of cDNA that was used as templates for method development.Through optimization by using three sets primers simultaneously,the detection limit of RNA template in the RT-PCR was 0.27 ng for CSFV,0.049 ng for PRRSV and 0.067 for JEV.No false positive results were produced in the case of transmissible gastroenteritis virus(TGEV),porcine parvovirus(PPV),pseudorabies virus(PrV) and porcine circovirus type 2(PCV-2).There were 174 samples,including sera,fetuses and lung tissues which were collected from the pigs that mainly presented high fever,were subjected to our in-house RT-PCR.PRRSV was detected in these samples with higher detection rate than CSFV and JEV.Among 174 clinical samples,PRRSV-positive percentage was 30.4,CSFV-positive percentage was 4.6,JEV-positive percentage was 1.7,co-infection of CSFV and PRRSV percentage was 1.7.In order to confirm the reliability of the multiplex PCR,we selected PCR products of CSFV-positive JEV-postive and PRRSV-positive respectively that detected clinical samples by multiplex PCR.After sequences analysis and BLAST,it is showed that homologies between PCR products of CSFV,PRRSV,JEV and target genes sequences were 92%,90% and 89%,respectively.Those results proved that multiplex RT-PCR have Applied value in practices." @default.
- W2394012938 created "2016-06-24" @default.
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- W2394012938 date "2008-01-01" @default.
- W2394012938 modified "2023-09-23" @default.
- W2394012938 title "Development of Multiplex RT-PCR to Detect Classical Swine Fever Virus(CSFV),Porcine Reproductive and Respiratory Syndrome Virus(PRRSV) and Japanese Encephalitis Virus(JEV)" @default.
- W2394012938 hasPublicationYear "2008" @default.
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