基于碳点标记的(1-3)-β-D葡聚糖的荧光检测方法

A High Selective and Specific Fluorescent Test Method for The Detection of (1,3)-β-D-Glucan

血清中(1,3)-β-D葡聚糖(βG)的含量已经成为临床上检测侵袭性真菌感染的指标,利用原核表达并纯化获得能够与βG特异性结合的美洲鲎属G因子α亚基片段a(Gαa)蛋白,将Gαa蛋白与碳点偶联获得生物荧光传感器"碳点/Gαa蛋白",建立基于碳点标记的βG荧光检测方法。经测定该传感器的最佳的激发波长为360nm,其发射波长为470nm,且荧光强度与βG浓度呈正线性相关,该检测方法线性范围是9.375~150pg/m L,最低检测限为9.375pg/m L,批间、批内精密度均不小于5%,准确度(回收率)在97%~102%之间,该方法灵敏、快速、成本低,具有广阔的应用前景。

The concentration of（1,3）-beta-D glucan（βG） in human serum is a clinical testing index of invasive fungal infection,a beta-glucan binding protein of horseshoe crab（limulus polyphemus） factor G-subunit α fragment-a（Gαa） was expressed in Escherichia coli BL21,Coupling of amination retouching carbon dots with G protein and establishing a＂carbon dots/Gαa protein＂fluorescent sensor for βG detection. The sensor excitation wavelength was 360 nm,emission wavelength was 460 nm.The fluorescent and the concentration of βG were a positive linear correlation.,the detection range was 9.375~150pg/m L,the limit of detection was 9.375 pg/m L. The CV values within batch and between batches were all less than 5%,accuracy（recovery） were between 97%~102%. The novel fluorescent sensor for the detection of βG was low cost,rapid,high selective,sensitivity and specificity.