MicroRNA通用探针法的建立及布鲁氏菌病患者血浆microRNA-146a的检测

发布时间：2018-05-19 00:39

本文选题：通用探针法 + 正交设计　；参考：《中国人兽共患病学报》2017年05期

【摘要】：目的建立一种通用、敏感、特异的microRNA检测方法,用于检测布鲁氏菌病患者血浆microRNA-146a的表达,探讨其作为诊断标识的可能性。方法以microRNA-146a模拟物为待检物,通过正交实验设计,对退火温度、探针浓度、试剂盒选择进行优化,建立特异敏感的通用检测方法;从布病患者和正常健康人血浆样本提取RNA,应用通用探针法进行检测,比较布病患者血浆microRNA-146a的表达变化。结果建立了优化的通用探针法,与染料法相比特异性更强、扩增范围更大,具有通用检测能力。应用通用探针法检测布鲁氏菌病患者血浆microRNA-146a,发现布病患者血浆中microRNA-146a的表达被抑制(P0.01),表明其可能作为布病诊断的分子标识。结论经优化的通用探针法是一种通用、敏感、特异的检测方法,可用于microRNA的检测,microRNA-146a可能作为布病诊断的标识分子。
[Abstract]:Objective to establish a universal, sensitive and specific microRNA assay for detection of plasma microRNA-146a expression in patients with brucellosis. Methods the microRNA-146a simulant was used as the test object. The annealing temperature, probe concentration and the selection of the kit were optimized by orthogonal design, and a specific and sensitive universal detection method was established. RNAs were extracted from the plasma samples of patients with brucellosis and healthy subjects. The expression of microRNA-146a in plasma of patients with brucellosis was detected by universal probe method. Results the optimized universal probe method was established. Compared with the dye method, the method was more specific, with a larger amplification range, and had the ability of universal detection. MicroRNA-146a in plasma of brucellosis patients was detected by universal probe method. It was found that the expression of microRNA-146a in the plasma of brucellosis patients was inhibited by P0.01a, indicating that it might be used as a molecular marker for the diagnosis of brucellosis. Conclusion the optimized universal probe method is a universal, sensitive and specific method for the detection of microRNA. MicroRNA-146a may be used as a marker for the diagnosis of brucellosis.
【作者单位】：军事兽医研究所军事医学科学院;沈阳农业大学;中国疾病预防控中心鼠疫布病防治基地;
【分类号】：R516.7

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