牛流行热病毒可视化RT-LAMP检测技术的建立及应用

发布时间：2018-06-14 18:29

  本文选题：牛流行热 + 病毒检测　； 参考：《华南农业大学学报》2017年03期

【摘要】：【目的】建立一种能够快速、简便、可视化地检测牛流行热病毒的分子生物学方法。【方法】根据牛流行热病毒G蛋白基因的6个保守区域设计2对引物,建立牛流行热病毒可视化逆转录环介导等温扩增(Reverse transcription loop-mediated isothermal amplification,RT-LAMP)检测技术,优化RT-LAMP的反应条件,将其与PCR方法进行比较。【结果】当Mg2+浓度为3 mmol·L~(-1)、甜菜碱浓度为0.4 mol·L~(-1)、d NTPs mix浓度为1.2μmol·L~(-1)、内外引物浓度比例为8∶1、反应温度为63℃时,反应梯形条带最明显,在反应40 min后可以观察到明显的梯形条带。建立的RT-LAMP检测方法特异性好,只对牛流行热病毒进行扩增;灵敏度比普通PCR高10倍。【结论】该方法操作简便,特异性强,结果判读方便,可用于牛流行热的快速检测。
[Abstract]:[objective] to establish a molecular biological method for rapid, simple and visual detection of bovine epidemic fever virus. [methods] two pairs of primers were designed according to 6 conserved regions of bovine prevalent fever virus G protein gene. In order to optimize the reaction conditions of RT-LAMP, a visualized reverse transcription loop-mediated isothermal amplification (RT-LAMPMPP) technique was established for the detection of bovine epidemic fever virus (BVV) by reverse transcription loop-mediated isothermal amplification. [results] when the concentration of Mg2 was 3 mmol / L ~ (-1), the concentration of betaine was 0.4 mol / L ~ (-1), the concentration of mix was 1.2 渭 mol / L ~ (-1), the ratio of internal and external primer was 8: 1 and the reaction temperature was 63 鈩，

本文编号：2018548