便携式血糖仪对耐甲氧西林金黄色葡萄球菌mecA基因的灵敏检测方法研究

摘要

本实验利用血糖仪构建了一种基于蔗糖酶修饰的纳米金功能化树状大分子对耐甲氧西林金黄色葡萄球菌mecA基因的灵敏检测新方法.当靶基因存在时,其与捕获核酸探针特异性结合,释放出带蔗糖酶的信号分子复合物,经磁分离后,上清液中的蔗糖酶催化底物蔗糖,产生可检测的葡萄糖信号,从而实现血糖仪对mecA基因的定量检测.该检测方法的线性范围是0.5 pM到1 nM,检测限为0.26 pM,优于一般的基因检测方法.结果表明,该方法在疾病诊断方面具有潜在的应用价值.%A sensitive quantitative detection method of mecA gene in methicillin-resistant Staphylococ-cus aureus based on nanogold-functionalized dendrimer using commercially available portable personal glu-cose meters(PGMs)was developed. In the presence of target gene,the specific hybridization occurred be-tween the capture probe and target gene,and the signal molecular complex with the invertase was re-leased. After separation by a magnetic rack,the invertase in the supernatant was used to catalyze the hy-drolysis of sucrose into glucose for PGM readout,establishing a bridge between PGMs and the quantitative detection of target gene. A linear relationship between the signals of PGMs and the concentrations of tar-get gene in the range of 0.5 pM to 1 nM was achieved. The detection limit was estimated at 0.26 pM, which was lower than the general gene detection methods. The results showed that this proposed method for sensitive detection of mecA gene using personal glucose meters had a potential application value in the disease diagnosis.