小鼠诺如病毒荧光定量PCR检测方法的建立及初步应用

发布时间：2018-03-07 12:08

本文选题：鼠诺如病毒MNV　切入点：实时荧光定量PCR　出处：《中国比较医学杂志》2016年12期 　论文类型：期刊论文

【摘要】：目的建立小鼠诺如病毒(murine norovirus,MNV)实时荧光定量PCR检测方法,为建立MNV规范化检测方法提供依据。方法根据NCBI公布的60个MNV病毒株核酸序列设计特异性引物,构建标准阳性质控品,建立MNV实时荧光定量PCR方法,并进行特异性、灵敏度、重复性及稳定性评价。用该方法对766只送检小鼠的盲肠内容物样品进行检测,初步了解北京地区实验小鼠MNV感染状况。结果成功建立MNV实时荧光定量PCR检测方法,该方法特异性好,与同种属人类诺如病毒(Hu No Vs)、猫杯状病毒(FCV)均无交叉反应,检测灵敏度可达101copies/μL。批内及批间CT值变异系数均小于2%。应用该方法在766份小鼠盲肠内容物样品中检出301份MNV核酸阳性样品,阳性率39.3%。结论建立的MNV实时荧光定量PCR检测方法特异性好,灵敏度高,重复性好,可以用于MNV的快速定量检测。
[Abstract]:Objective to establish a real-time fluorescent quantitative PCR method for the detection of murine norovirusus of mouse norovirus.Methods according to the nucleic acid sequences of 60 MNV virus strains published by NCBI, specific primers were designed and the standard positive quality control products were constructed. MNV real-time fluorescence quantitative PCR method was established to evaluate the specificity, sensitivity, reproducibility and stability of cecum contents in 766 mice. Results the real-time fluorescence quantitative PCR assay for the detection of MNV was successfully established. The method was specific and had no cross reaction with Hu No VsSN of the same species and calicivirus of cat. The sensitivity of detection was 101 copias / 渭 L. the coefficient of variation of CT values in and between batches was less than 2. 301 MNV nucleic acid positive samples were detected in 766 mouse caecum contents by this method. Conclusion the established MNV real-time fluorescent quantitative PCR method has good specificity, high sensitivity and good reproducibility, and can be used for rapid quantitative detection of MNV.
【作者单位】：中国食品药品检定研究院实验动物资源研究所;
【基金】：国家科技支撑课题(2013BAK11B01)
【分类号】：R440

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