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发明授权
CN101914631B 小鼠肝炎病毒实时荧光定量PCR检测方法 有权 转让 许可

小鼠肝炎病毒实时荧光定量PCR检测方法
摘要：
本发明涉及一种小鼠肝炎病毒实时荧光定量PCR检测方法。该方法包括标准质粒的构建、特异性引物及荧光探针的设计、实时荧光定量PCR扩增及建立检测标准曲线、感染病毒样本的RNA提取及cDNA的制备以及有效性验证及结果检测判定步骤；其中特异性引物的正向引物的序列为：5’-CCCGAAGTGGTTAGTGAAGG-3’，反向引物的序列为：5’-CATGGCCCTATAATACGGGT-3’；所述荧光探针的序列为：5’-CTCGCTCTATGACCTACTGC-3’。本发明检测灵敏度高，检测时间短，可同时进行大批量的样本分析，具有良好的特异性、敏感性、重复性和稳定性，适用于病毒感染的前期诊断。
摘要（英）：

The invention relates to a real-time fluorescent quantitative PCR detection method for mouse hepatitis virus. The method comprises the following steps of: constructing a standard plasmid, designing a specific primer and a fluorescent probe, performing real-time fluorescent quantitative PCR amplification, establishing a detection standard curve, extracting RNA from a virus infected sample, preparing cDNA, verifying effectiveness and detecting and judging a result, wherein the sequence of a forward primer of the specific primer is 5'-CCCGAAGTGGTTAGTGAAGG-3', while the sequence of a reverse primer is 5'-CATGGCCCTATAATACGGGT-3'; and the sequence of the fluorescent probe is 5'-CTCGCTCTATGACCTACTGC-3'. The method has the advantages of high detection sensitivity, short detection time, simultaneous mass sample analysis, high specificity, sensitivity, repeatability and stability, and suitability for early diagnosis of virus infection.

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