【摘要】：為實(shí)現(xiàn)對(duì)鴨圓環(huán)病毒(DuCV)的快速檢測(cè),分析比對(duì)Gen Bank中登錄的DuCV全基因組序列,在其保守區(qū)設(shè)計(jì)并合成一對(duì)特異性引物,將擴(kuò)增的212 bp片段克隆入pMD-18T載體中,獲得pMD18DuCV重組質(zhì)粒,通過(guò)SYBR GreenⅠ熒光摻入法分別對(duì)系列稀釋pMD18DuCV中DuCV特異性片段進(jìn)行擴(kuò)增,通過(guò)pMD18DuCV濃度對(duì)數(shù)與Ct值制作了標(biāo)準(zhǔn)曲線(xiàn)并推導(dǎo)出直線(xiàn)回歸方程,二者相關(guān)系數(shù)(R~2)為0.9968,建立了DuCV實(shí)時(shí)熒光定量PCR檢測(cè)方法,重復(fù)性和靈敏性檢測(cè)結(jié)果表明重復(fù)性良好,檢測(cè)下限為67.2拷貝/μL;將建立的DuCV實(shí)時(shí)熒光定量PCR檢測(cè)方法分別對(duì)鴨肝炎病毒(DHV)、大腸桿菌、新城疫病毒(NDV)、H9亞型禽流感病毒、小鵝瘟病毒(GPV)核酸或cDNA進(jìn)行特異性檢測(cè),結(jié)果表明特異性良好。初步應(yīng)用結(jié)果表明,建立的DuCV實(shí)時(shí)熒光定量PCR方法檢出率為26.4%(62/235),明顯優(yōu)于常規(guī)PCR檢測(cè)結(jié)果15.3%(36/235)。
[Abstract]:In order to detect duck circovirus (DuCV) quickly, a pair of specific primers were designed and synthesized in the conserved region of Gen Bank to analyze and compare the whole DuCV genome sequence registered in Gen Bank. The amplified 212bp fragment was cloned into pMD-18T vector to obtain pMD18DuCV recombinant plasmid. The specific fragments of DuCV in diluted pMD18DuCV were amplified by SYBR Green 鈪，

本文編號(hào)：2192267