犬SLAM受体mRNA荧光定量PCR检测方法的建立和初步应用  

Development and application of fluorescence quantitative RT-PCR for detection of slam mRNA in canine tissues

作　　者：夏菁[1] 李佳[2] 姜代勋[2] 陈武[1] 

机构地区：[1]北京农学院,动物科学科技学院,北京102206 [2]兽医学(中医药)北京市重点实验室,北京102206

出　　处：《中国比较医学杂志》2013年第6期44-48,共5页Chinese Journal of Comparative Medicine

基　　金：北京市属高等学校人才强教计划资助项目(PHR201107134)

摘　　要：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。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 0. 89% - 2. 35%. The RT-PCB assay had advantages of high reproducibility. The SLAM mRNA was transeripted 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 mBNA transcription of SLAM in different canine tissues.

关 键 词：犬 SLAM受体 mRNA转录水平 荧光定量PCR 

分 类 号：R33[医药卫生—人体生理学]