鼠源炎症因子流式蛋白定量试剂盒的优化及初步应用

Optimization of Mouse Inflammation Cytometric Bead Array Kit by Flow Cytometry and Its Application

以小鼠炎症因子流式蛋白定量(cytometric bead array,CBA)检测试剂盒为参考,分别从降低试剂用量和缩短孵育时间两个方面进行优化,然后以沙门菌感染小鼠为模型,动态分析小鼠血清中炎症因子的分泌情况。结果表明,试剂用量由50μL降至20μL、孵育时间由2h缩短至30min仍然可以得到较好的检测效果。沙门菌感染小鼠后,利用该方法动态分析了小鼠血清中炎症因子的变化情况,并且发现炎症因子的早期分泌与沙门菌在小鼠体内的定殖呈正相关。通过CBA技术的优化和初步应用,为其推广应用提供了重要数据。

The mouse inflammation cytometric bead array （CBA） kit was used as reference,the mean fluo- rescence intensity by flow cytometry was compared through changing reagent dose and incubation time,respectively.Then the serum cytokines of mice were measured after infection with Salmonella.The results indicated that similar tendency can be detected when decreasing reagent dose from 50 μL to 20μL and shortening incubation time from 2 hours to 30 minutes, respectively. The optimized CBA technique was then used to analyze the serum cytokines in Salmonella-infected mice.We simultaneously found that the secre- tion of inflammatory cytokines in the early-stage of infection is associated with the colonization of Salmo- nella in mice.This data will be helpful for the application of CBA technique.