建立一种使用小沟结合探针的一步法实时 RT-PCR 检测方法用于鸭坦布苏病毒的检测。
Development of a one-step real-time RT-PCR assay using a minor-groove-binding probe for the detection of duck Tembusu virus.
机构信息
Institute of Animal Husbandry and Veterinary Sciences, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China.
出版信息
J Virol Methods. 2012 May;181(2):148-54. doi: 10.1016/j.jviromet.2012.01.019. Epub 2012 Feb 2.
The design and development of a 3'-conjugated minor-groove-binding (MGB) probe for a real-time RT-PCR assay allowing for the rapid, sensitive, and specific detection of duck Tembusu virus (DTMUV) RNA are described. This assay targeted the 3' terminal non-coding region (NCR) of the TMUV genome and detected 1 × 10¹ copies of RNA per reaction without cross-reaction with other duck pathogens. The linear range of detection was 2 × 10¹-2 × 10⁸ copies/μl. The assay was rapid, requiring just over 2.0 h, including the nucleic acid extraction step. Therefore, this assay is an excellent tool for research routine diagnostic applications, and study of the epidemiology of TMUV infections among duck flocks.
描述了一种用于实时 RT-PCR 检测的 3'-共轭小沟结合（MGB）探针的设计和开发，该探针能够快速、敏感、特异地检测鸭坦布苏病毒（DTMUV）RNA。该检测方法针对 TMUV 基因组的 3'末端非编码区（NCR），每个反应可检测到 1×10¹拷贝的 RNA，与其他鸭病原体无交叉反应。检测的线性范围为 2×10¹-2×10⁸拷贝/μl。该检测方法快速，仅需 2.0 小时以上，包括核酸提取步骤。因此，该检测方法是研究常规诊断应用和鸭群中 TMUV 感染流行病学的优秀工具。
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