【摘要】 将改良的实时TaqMan荧光定量RT-PCR技术应用于口蹄疫病毒感染体内和体外的定量检测以及其3D基因转录水平分析。结果表明:对样品中口蹄疫病毒基因组RNA的检测灵敏度可达l0个基因拷贝,可同时检测病毒正负链复制水平且重复性较好,所测口蹄疫病毒3D基因转录水平可高达6.9×104拷贝/μL;与实时SYBRGreenⅠ染料RT-PCR技术比较,改良的实时TagMan荧光定量RT-PCR技术检测灵敏度高6.7倍。以上结果证实,改良的实时TaqMan荧光定量RT-PCR技术在病毒检测和基因表达水平分析方面有更高的灵敏度和特异性。更多还原

【Abstract】 In this study, an improved real-time RT-PCR was applied to quantification of Foot-and-mouth disease virus in vivo and in vitro. The results suggested that this method had a high sensitivity with up to less than 10 copies / reaction. Furthermore, this novel method was successfully used to quantify the transcriptional level of FMDV-3D gene in transfected BHK-21 cells and the resultant concentration of RNA level was up to 6.9×10 4 copies/μL. Compared with SYBR GreenⅠRT-PCR, six of eight samples testing the FMDV RNA copy number was up to 6.7-fold higher when determined by TaqMan RT-PCR, indicating that the improved method shows a high sensitivity in virus detection.更多还原