犬SLAM受体mRNA荧光定量PCR检测方法的建立和初步应用

摘要

Objective To establish a method for the detection of mRNA transcription of SLAM as CDV receptors in different canine tissues.Methods By using GAPDH as a housekeeper,The △△ Ct relative quantification method was used to detect the transcription levels of SLAM mRNAs in different canine tissues.Results The inter-assay variability (CV％) was O.89％-2.35％.The RT-PCR assay had advantages of high reproducibility.The SLAM mRNA was transcripted at high levels in the spleen,hilar lymph node,mesenteric nodes,inguinal nodes and a low level in the urinary bladder.Conclusions The quantitative RT-PCR method is successfully established for the detection of mRNA transcription of SLAM in different canine tissues.%目的 建立荧光定量PCR方法,检测犬不同组织中SLAM受体mRNA的表达水平.方法 以犬GAPDH为内参基因采用△△Ct法,分析SLAM受体mRNA在犬体内不同组织中的表达.结果 此方法有较高的重复性,变异系数在0.89％～2.35％.以SLAM受体在心脏的表达为1倍值,结果显示受体mRNA在脾脏中表达最高,为38.49倍；肺门淋巴结、肠系膜淋巴结、腹股沟淋巴结中表达次之,分别为9.13、8.58、6.24倍；膀胱中表达最低.结论 成功建立了检测SLAM受体mRNA在不同组织中表达水平的荧光定量PCR检测方法.