树鼩感染相关细胞因子荧光定量PCR检测方法的建立  

Construction of real-time polymerase chain reaction detection for infection-related cytokines of tree shrew

作　　者：李潇[1] 刘文宽[1] 邱淑燕[1] 许多[1] 周志超[1] 田新贵[1] 李炽 辜淑君 周荣[1] LI Xiao;LIU Wenkuan;QIU Shuyan;XU Duo;ZHOU Zhichao;TIAN Xingui;LI Chi;GU Shujun;ZHOU Rong(State Key Laboratory of Respiratory Diseases,National Clinical Research Center forRespiratory Disease,Guangzhou Institute of Respiratory Health,The First Affiliated Hospital of Guangzhou Medical University,Guangzhou MedicalUniversity,Guangzhou 510120,P.R.China)

机构地区：[1]呼吸疾病国家重点实验室国家呼吸系统疾病临床医学研究中心广州医科大学广州医科大学附属第一医院广州呼吸健康研究院

出　　处：《生物医学工程学杂志》2019年第3期407-413,共7页Journal of Biomedical Engineering

基　　金：国家自然科学基金(31500143);国家科技重大专项(2018ZX10102001,2017ZX10103011);广东省自然科学基金项目(2016A030313572);广州市科技计划项目(201803040004)

摘　　要：树鼩是一种新型的优质实验动物模型。本研究针对树鼩感染相关的细胞因子白介素6(IL-6)、IL-8、IL-10、IL-17A、γ-干扰素(IFN-γ)及管家基因磷酸甘油醛脱氢酶(GAPDH)建立荧光定量聚合酶链式反应检测方法。研究证明所建立方法具有良好的特异性。线性范围高点可达到1×1010copies,低点则至10copies(IL-6、IL-17A)、100copies(IL-10、GAPDH)和1000copies(IL-8、IFN-γ)不等,在此区间各试剂线性相关系数R2均大于0.99。IL-6、IL-8、IL-10、IL-17A、IFN-γ和GAPDH最低检测值分别在8、8、4、8、128、4copies。研究表明所建立的检测方法具有很好的特异性、灵敏度及较广的线性范围,适用于多重浓度范围的样品检测,可用于后续树鼩细胞因子的研究工作。Tree shrew is a novel and high-quality experimental animal model.In this study,the real-time polymerase chain reaction methods were established to detect infection-related cytokines interleukin-6(IL-6),IL-8,IL-10,IL-17A,interferon-γ(IFN-γ)and housekeeping gene glyceraldehyde-phosphate dehydrogenase(GAPDH)of tree shrew.The results indicated that the establised methods had good specificity.The high point of the linear range of these reagents reached 1×1010 copies,and the low points ranged from 10 copies(IL-6,IL-17A),100 copies(IL-10,GAPDH)to 1 000 copies(IL-8,IFN-γ).In this interval,the linear correlation coefficient R2 of each reagent was greater than 0.99.The lowest detectable values of IL-6,IL-8,IL-10,IL-17A,IFN-γand GAPDH were 8,8,4,8,128 and 4 copies,respectively.The results showed that the established detection methods had good specificity,sensitivity and wide linear range.The methods were suitable for detection of multiple concentration range samples,and could be used for the subsequent studies of tree shrew cytokines.

关 键 词：树鼩 细胞因子 荧光定量聚合酶链式反应 

分 类 号：R-332[医药卫生]