CAJ | 학술논문

本研究依据布鲁氏菌的特异性基因Omp25c的部分片段作为靶基因，设计探针引物，且优化了反应体系，筛选出引物、探针的最优浓度配比。将扩增产物连接到PGEM-T载体上，制备标准品及标准曲线，建立鹿布鲁氏菌荧光定量PCR检测方法，并对其特异性、稳定性、敏感性进行评价。由标准曲线可知该方法的最低检测浓度可达到36拷贝/μL，比常规PCR灵敏度高出很多。
본연구의거포로씨균적특이성기인Omp25c적부분편단작위파기인，설계탐침인물，차우화료반응체계，사선출인물、탐침적최우농도배비。장확증산물련접도PGEM-T재체상，제비표준품급표준곡선，건립록포로씨균형광정량PCR검측방법，병대기특이성、은정성、민감성진행평개。유표준곡선가지해방법적최저검측농도가체도36고패/μL，비상규PCR령민도고출흔다。
To aid in the development of a rapid assay to detect Brucella in deer,specific probe and primers was optimized based on the conservative Omp25c segment of Brucella and the reaction system was improved for selecting the best concentration of the primers and the probe. The amplified gene was connected to PGEM-T vector,and the developed RT-PCR assay was evaluated with regard to its specificity,replicability and sensitivity via constructing standard samples and standard curve. The result showed that the detecting limit was 36 copies/μL and more sensitive than the general PCR.