A highly sensitive and selective DNA biosensor is described based on the fluorescence quenching ability of MoS2 nanosheet and exonucleaseⅢ( ExoⅢ) assisted dual-signal amplification. In this sensor, the fluorescence probes ( HP1 and HP2 ) cannot be degraded by Exo Ⅲdue to the 3 '-termini protrusion and thus are adsorbed on the surface of MoS2 nanosheets, which will result in the quenching of MoS2 nanosheets toward the probes and induce a low fluorescent signal. The presence of the target DNA leads to the desorption of probes on the surface of MoS2 nanosheets due to the hybridization toward probes, generating many fluorescent fragments by Exo Ⅲdigestion and inducing dual-signal amplification. This method can improve the sensitivity and detection limit compared with single amplification method, and shows excellent selectivity in the discrimination of single base mismatched targets. On the basis of the significantly high sensitivity, the developed biosensor can be potentially extended to detect various DNA targets with excellent sequence selectivity.%将核酸外切酶Ⅲ诱导的双重信号放大技术与MoS2纳米片的荧光猝灭性质结合,构建了一种高灵敏高选择性的DNA检测方法.首先设计两条末端修饰荧光基团的探针核酸(HP1和HP2).由于两条探针核酸具有3'粘性末端,使其不会被核酸外切酶Ⅲ降解,因而被吸附于MoS2纳米片而猝灭其荧光.当目标DNA存在时,会促使核酸外切酶Ⅲ启动双重信号放大反应,并将探针核酸降解成大量的不能吸附于MoS2纳米片表面的荧光碎片.在优化条件下,目标DNA浓度在0.5~6.0 pmol/L范围内与荧光信号变化呈良好的线性关系,检出限为0.28 pmol/L.与单重信号放大技术相比,本方法极大改善了分析灵敏度和检出限,且具有良好的单碱基错配区分能力.
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