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中文摘要：
根据鸭坦布苏病毒（DuckTembusuvirus，DFV）NS5基因序列特征设计引物，建立基于SYBRGreenI检测模式的实时荧光定量RT—PCR（real—timeRT—PCR。RRT—PCR），该方法检测DFVNS5基因2．74×10。～2．74×10’拷贝／肛L反应范围内有很好的线性关系。扩增产物的熔解曲线分析只出现1个单特异峰，无引物二聚体，丁m值为（86．23士0．18）。C，对禽流感病毒、鸭肝炎病毒、鸭源禽1型副黏病毒、鸭减蛋综合征病毒、番鸭呼肠孤病毒核酸均无阳性信号扩增，可重复性好，组内变异系数为0．52％～1．48％，组间变异系数0．71％～2．21％。检测速度快，从样本处理到报告结果仅需4h。
英文摘要：
A pair of specific primers targeted to non structure gene 5 （NSS） of duck Tembusu virus was designed and a SYBR Green I fluorescent based real-time RT-PCR （RRT-PCR） was devel- oped for the quantization of duck Tembusu virus. The detection limit of RRT-PCR was 2.74 × 102 plasmid copies. The melting curve analysis using SYBR Green I dye showed one specific peak, a melting temperature （Tm） was （86.23±0.18） ℃ ,and no primer-dimers peak was observed. No amplification was detected from unrelated virus samples by this method, such as avian influenza virus, duck hepatitis virus type 1, avian paramyxovirus type 1, egg drop syndrome virus, duck reo- virus. Fine reproducibility was obtained for detecting plasmid DNA with intra-assay of 0. 52%- 1.48G and inter-assay of 0.71G-2.21G. The real-time PCR method developed in this study will be useful for rapid laboratory diagnosis and epidemiology investigation for duck Tembusu virus.
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