發(fā)布時間：2018-03-28 22:26

  本文選題：犬圓環(huán)病毒　切入點：SYBR　出處：《動物醫(yī)學(xué)進展》2017年05期

【摘要】：犬圓環(huán)病毒(DogCV)是圓環(huán)病毒科新發(fā)現(xiàn)的圓環(huán)病毒,旨在建立犬圓環(huán)病毒實時熒光定量PCR檢測方法。試驗用PCR方法克隆犬圓環(huán)病毒ORF2基因保守區(qū)域,構(gòu)建標(biāo)準(zhǔn)陽性質(zhì)粒pClon007-ORF2。以標(biāo)準(zhǔn)陽性質(zhì)粒pClon007-ORF2為模板對熒光定量PCR反應(yīng)體系進行優(yōu)化。結(jié)果表明,所建立的方法 Ct值與標(biāo)準(zhǔn)品在4.67×10~9 copies/μL~4.67×10~3 copies/μL范圍內(nèi)呈良好的線性關(guān)系,相關(guān)系數(shù)為0.998,斜率為-3.445。該方法檢測靈敏度為4.67×10~1copies/μL。該方法對狂犬病病毒、犬細(xì)小病毒、犬瘟熱病毒、犬副流感病毒、犬腺病毒2型等犬常見病原的檢測均無交叉反應(yīng);所有稀釋度標(biāo)準(zhǔn)品模板均在83.35℃出現(xiàn)窄的特異性熔解峰。臨床樣品檢測表明,所建立的實時熒光定量PCR對犬圓環(huán)病毒的陽性檢測率為12.50%(4/32)。
[Abstract]:Dog circovirus (DogCVV) is a newly discovered circovirus in the family of circoviridae. The aim of this study was to establish a real-time fluorescent quantitative PCR assay for the detection of canine circovirus. The conserved region of ORF2 gene of DogCVV was cloned by PCR method. The standard positive plasmid pClon007-ORF2 was constructed. The fluorescent quantitative PCR reaction system was optimized by using standard positive plasmid pClon007-ORF2 as template. The results showed that the established method had a good linear relationship with the standard sample in the range of 4.67 脳 10 ~ (9) copies/ 渭 L ~ (4.67) 脳 10 ~ (-3) copies/ 渭 L. The correlation coefficient was 0.998, the slope was -3.4455.The sensitivity of this method was 4.67 脳 10~1copies/ 渭 L. there was no cross reaction in the detection of rabies virus, canine parvovirus, canine distemper virus, canine parainfluenza virus and canine adenovirus type 2. All the dilution standard sample templates showed narrow specific melting peak at 83.35 鈩，

本文編號：1678358