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中文摘要：
为建立鸭疫里默氏杆菌（Riemerella anatipestife，RA）定量检测方法，本研究根据RA的铁依赖抑制子基因（RaDtxR）序列，设计特异性引物和探针，建立基于TaqMan探针检测RA的荧光定量PCR方法。该方法检测RA RaDtxR在4.27×10^2拷贝/μL～4.27×10^6拷贝/μL范围内具有良好的线性关系，其扩增相关系数为0.998，扩增效率为99 %，最低检测下限为4.27×10^2拷贝/μL。对常见RA血清型（1型、2型、3型和10型）检测结果均为阳性；而对大肠杆菌、巴氏杆菌、沙门氏菌、Ⅰ型鸭肝炎病毒、禽坦布苏病毒和鸭圆环病毒等常见鸭群病原检测均为阴性。该方法重复性好，组内和组间变异系数分别为0.23 %～0.77 %和0.93 %～1.32 %。该方法的建立为RA感染宿主靶器官的感染程度和定量分析奠定基础。
英文摘要：
A TaqMan-based real-time PCR was developed for detection of Riemerella anatipestifer with the primers and probe targeting the RaDtxR gene. The standard curve established with the positive plasmid had a wide dynamic range from 4.27×10^2 copies/μL to 4.27×10^6 DNA copies/μL of R.anatipestifer with a linear correlation （R2） of 0.998 and efficiency of 99% between the Ct value and the logarithm of the plasmid copy number. No amplification was detected from other bacteria or virus, such as E.coli, Pasteurella, Salmonella, duck hepatitis virus, avian Tembusu virus and duck circovirus. Reproducibility test showed that the intra- and inter-assay were 0.23% to 0.77% and 0.93% to 1.32%, respectively. The establishment of the real-time PCR method in this study would be useful for earlier diagnosis and quantitative detection of R.anatipestifer load in organs of infected duck.
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