使用多探针核糖核酸酶保护分析同时定量马细胞因子信使核糖核酸。
Simultaneous quantitation of equine cytokine mRNAs using a multi-probe ribonuclease protection assay.
作者信息
Lim W-S, Edwards J F, Boyd N K, Payne S L, Ball J M
机构信息
Department of Veterinary Pathobiology, Texas A&M University, MS 4467, College Station, TX 77843-4467, USA.
出版信息
Vet Immunol Immunopathol. 2003 Jan 10;91(1):45-51. doi: 10.1016/s0165-2427(02)00263-5.
A rapid multi-probe ribonuclease protection assay (RPA) was developed to quantitate equine-specific cytokine mRNA levels in activated equine monocyte-derived macrophages (EMDM) and equine peripheral blood mononuclear cells (EPBMC). Eleven template plasmids specific to 10 equine cytokine genes and the beta-actin gene were generated from which radiolabeled anti-sense RNA probes were produced. The multi-probe RPA simultaneously quantitated mRNA levels of equine IL-1alpha, IL-1beta, IL-6, IL-8, IL-10, IL-12 p35, IL-12 p40, IFN-gamma, TGF-1beta and TNF-alpha in EPBMC and EMDM with coefficients of variation as low as 0.03-0.08 (3-8%) when normalized to beta-actin expression. This sensitive and rapid assay provides a valuable tool for studies of equine immune responses.
开发了一种快速多探针核糖核酸酶保护分析（RPA）方法，用于定量活化的马单核细胞衍生巨噬细胞（EMDM）和马外周血单个核细胞（EPBMC）中马特异性细胞因子mRNA水平。针对10种马细胞因子基因和β-肌动蛋白基因生成了11种模板质粒，从中制备放射性标记的反义RNA探针。多探针RPA同时定量EPBMC和EMDM中马IL-1α、IL-1β、IL-6、IL-8、IL-10、IL-12 p35、IL-12 p40、IFN-γ、TGF-1β和TNF-α的mRNA水平，以β-肌动蛋白表达进行标准化时，变异系数低至0.03-0.08（3-8%）。这种灵敏且快速的分析方法为马免疫反应的研究提供了一种有价值的工具。
Zotero 插件