新型核酸酶传感器测定血铅

Establishment of a Novel DNAzyme Sensor for Detecting Lead in Blood

研究建立基于核酸酶传感器（铅离子特异性DNA酶）的血铅检测体系。将全血标本依次经Triton X-100、蛋白酶K、稀硝酸及三氯乙酸处理,校正酸碱度后与核酸酶传感器混合进样,读取荧光增长速率进行定量检测。以标准铅溶液制作标准曲线,测定血铅标准物质以评价精密度及回收率。结果显示,线性范围良好（0~2000nmol/L,R2=0.9988）,方法检出限（LOD）9.44nmol/L。血铅标准物质检测的准确度及精密度良好（RSD分别为4.99%和1.35%）,方法回收率90.7%~95.8%。本实验成功建立了新型核酸酶传感器测定血铅的样本前处理方法及检测体系,该方法可以快速、准确地对血铅进行定量分析,为临床诊断提供帮助。

To develop a method for detecting lead in blood by a novel DNAzyme sensor（Pb dependent DNAzyme）.Blood samples were pretreated by Triton X-100,Proteinase K,nitric acid and TCA in turn.After adjusting pH to neutral,pretreated blood sample was mixed with substrate strand and enzyme strand.Time-dependent fluorescent measurement at ex/em= 485/535 nm was immediately started.The rate of fluorescent enhancement was recorded and analyzed with concentration of lead in the sample.Calibration curve was linear to 2000nmol/L,with R2=0.9988.Limit of detection was 9.44nmol/L.Detection of two reference materials of blood lead showed ideal accuracy and imprecision.（RSD of the two reference materials were 4.99% and 1.35%）.In four recovery studies,recoveries were target.A novel DNAzyme sensor for detecting lead in blood has been developed,which could be used in clinical detection.