CAJ | 학술논문

针对火鸡疱疹病毒（HVT ）特异性的 sorf 1区基因序列设计引物，建立检测 HVT 的SYBR Green Ⅰ实时荧光定量PCR方法，并对其特异性、敏感性和重复性试验验证。结果表明，建立的实时荧光定量 PC R方法标准曲线的循环阈值（C t ）与模版拷贝数呈良好线性关系（r2＝0．99），熔解曲线分析显示该PC R扩增具有良好的特异性、敏感性和重复性，试验表明该方法最低检测浓度为7×101拷贝／μL。利用该方法对野生型 HVT和rHVT‐pmpD‐N在体内和体外的复制特性检测，结果表明重组后的HVT与野生型HVT的增殖速度基本一致。本试验建立的检测方法能够快速检测 HV T ，准确率高，特异性好，具有较高的灵敏度和稳定性，为监测HV T体内和体外的病毒含量和复制情况提供了一种有效的手段。
침대화계포진병독（HVT ）특이성적 sorf 1구기인서렬설계인물，건립검측 HVT 적SYBR Green Ⅰ실시형광정량PCR방법，병대기특이성、민감성화중복성시험험증。결과표명，건립적실시형광정량 PC R방법표준곡선적순배역치（C t ）여모판고패수정량호선성관계（r2＝0．99），용해곡선분석현시해PC R확증구유량호적특이성、민감성화중복성，시험표명해방법최저검측농도위7×101고패／μL。이용해방법대야생형 HVT화rHVT‐pmpD‐N재체내화체외적복제특성검측，결과표명중조후적HVT여야생형HVT적증식속도기본일치。본시험건립적검측방법능구쾌속검측 HV T ，준학솔고，특이성호，구유교고적령민도화은정성，위감측HV T체내화체외적병독함량화복제정황제공료일충유효적수단。
The purpose of this study was to develop a real‐time PCR assay to detect and quantify herpesvir‐us of turkey(HVT)in vivo and in vitro targeting the unique sorf 1 gene of the virus .This assay was carried out using a LingtCycler instrument and the product was monitored constinuously with the fluorescent double‐stranded DNA binding dye SYBR Green Ⅰ .The duplicate character of wild HVT and recombinant HVT‐pmpD‐N(rHVT‐pmpD‐N)was detected using this method .Standard curve was established and the correlation index was 0 .99 .The result showed that SYBR Green Ⅰ real‐time PCR had a minimum detec‐tion limit of 7 × 101 copies/μL which was 10 times higher in sensitivity than standard PCR .SYBR Green Ⅰreal‐time PCR assay also showed a similar replication rate between wild HVT and rHVT‐pmpD‐N .It was concluded that this could be an effective ,economic and reliable method for rapid detection of wild HVT and recombinant HVT .