CAJ | 학술논문

为了建立检测(番鸭)呼肠孤病毒(DRV)感染的方法,本研究根据DRV的σNS基因核苷酸序列设计引物及TaqMan探针,经过反应条件的优化,建立了该病毒的TaqMan探针实时荧光RT-PCR检测方法.用已知浓度的重组质粒为标准品,构建的标准曲线的相关系数达到99%.试验表明,该方法的重复性、稳定性、特异性良好,且灵敏度高,可以检测到1.0×10~2拷贝/μL的标准品,比普通PCR至少高100倍.该检测方法对DRV人工感染番鸭肝脏组织检出率达到100%.本实验所建立的TaqMan探针荧光RT-PCR为的快速检测以及进行分子流行病学的调查提供了新的技术手段.
위료건립검측(번압)호장고병독(DRV)감염적방법,본연구근거DRV적σNS기인핵감산서렬설계인물급TaqMan탐침,경과반응조건적우화,건립료해병독적TaqMan탐침실시형광RT-PCR검측방법.용이지농도적중조질립위표준품,구건적표준곡선적상관계수체도99%.시험표명,해방법적중복성、은정성、특이성량호,차령민도고,가이검측도1.0×10~2고패/μL적표준품,비보통PCR지소고100배.해검측방법대DRV인공감염번압간장조직검출솔체도100%.본실험소건립적TaqMan탐침형광RT-PCR위적쾌속검측이급진행분자류행병학적조사제공료신적기술수단.
A real-time PCR assay was developed for detection of duck reovirus (DRV) infection using specific primers and TaqMan probes based on the σNS gene of DRV. The assay could detect 1.0 × 10~2 copies/μL of the standards, which is more than 100-fold sensitive than conventional PCR. This method was used to detect DRV replication in the liver of infected muscovy ducks and achieved 100 % success rate. Taken together, the TaqMan probe RT-PCR provides an effective method for rapid and accurate detection of DRV and for molecular epidemiology study of the virus infection.