摘要
为寻求喀斯特炭疽菌的快速检测方法,利用TaqMan探针实时荧光定量PCR技术,以喀斯特炭疽菌基因组中的ACT基因为靶序列设计并筛选特异性引物探针,通过特异性、灵敏度、重复性试验建立喀斯特炭疽菌TaqMan探针实时荧光PCR检测方法。结果表明:建立方法的特异性较强、灵敏度较高、重复性稳定性较好,最低检测限为0.2pg DNA/反应,标准曲线的线性关系良好,相关系数为0.998。该方法操作简便,可用于进出境口岸喀斯特炭疽菌的检疫。
The TaqMan real-time fluorescence PCR detection method of Colletotrichum karstii was established by using the TaqMan real-time fluorescence quantitative PCR technology, taking ACT gene as the target and screening specific primers and probe after specificity, sensitivity and repetition tests to explore the rapid detection method of Colletotrichum karstii. Results： The minimum detection limit of the established TaqMan real-time fluorescence PCR detection method with strong specificity, high sensitivity, stable repeatability, good linear relation （0. 998） and simple operation is 0.2 pg DNA, which indicates the established TaqMan real-time fluorescence PCR detection method can be used in Colletotrichum karstii; quarantine at entry-exit ports.
出处
《贵州农业科学》
CAS
2017年第2期5-9,共5页
Guizhou Agricultural Sciences
基金
国家质量监督检验检疫总局科技计划项目"基于DNA条形码的三种重要炭疽菌快速检测技术研究"(2015IK336)
