本文選題：通用探針法 + 正交設(shè)計　； 參考：《中國人獸共患病學報》2017年05期

【摘要】：目的建立一種通用、敏感、特異的microRNA檢測方法,用于檢測布魯氏菌病患者血漿microRNA-146a的表達,探討其作為診斷標識的可能性。方法以microRNA-146a模擬物為待檢物,通過正交實驗設(shè)計,對退火溫度、探針濃度、試劑盒選擇進行優(yōu)化,建立特異敏感的通用檢測方法;從布病患者和正常健康人血漿樣本提取RNA,應(yīng)用通用探針法進行檢測,比較布病患者血漿microRNA-146a的表達變化。結(jié)果建立了優(yōu)化的通用探針法,與染料法相比特異性更強、擴增范圍更大,具有通用檢測能力。應(yīng)用通用探針法檢測布魯氏菌病患者血漿microRNA-146a,發(fā)現(xiàn)布病患者血漿中microRNA-146a的表達被抑制(P0.01),表明其可能作為布病診斷的分子標識。結(jié)論經(jīng)優(yōu)化的通用探針法是一種通用、敏感、特異的檢測方法,可用于microRNA的檢測,microRNA-146a可能作為布病診斷的標識分子。
[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.
【作者單位】： 軍事獸醫(yī)研究所軍事醫(yī)學科學院;沈陽農(nóng)業(yè)大學;中國疾病預(yù)防控中心鼠疫布病防治基地;
【分類號】：R516.7

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