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用基于生物传感器的快速尿检检测泌尿生殖道血吸虫病

By LabMedica International staff writers
Posted on 13 Oct 2015
在基础设施薄弱的地区,护理点(POC)分子诊断有望变革血吸虫病等传染病的诊疗,这些传染病如果得不到诊断,人会长期带病。

因为电化学生物传感器灵敏度高、成本低,容易集成进POC设备,兼之酶标仪小巧便携,所以非常适合分子诊断,现在已经用于检测泌尿生殖道血吸虫病。

美国加利福尼亚州斯坦福大学医学院的科学家及其同事开发的方法可用电化学生物传感器在一小时内对尿道细菌感染进行分子诊断。尿细胞被裂解,再直接涂到一排传感器上,传感器事先用靶向常见尿路病原体的16S核糖体核糖核酸(rRNA)的寡核苷酸探针进行过功能化。病原体rRNA和标记过的捕获探针和检测探针对之间形成序列特异的杂交复合体,一个介导电流信号输出的酶标签可检测该复合体。

生物传感器由三个平面金电极组成,一个工作电极,一个辅助电极,一个参考电极。生物传感器测定产品的捕获探针通过巯基连接到工作电极表面。样本里的细胞被裂解并与检测探针的缓冲溶液混合,然后涂到传感器表面。如果目标rRNA存在,就会和捕获探针与检测探针一起形成杂交复合体。辣根过氧化物酶(HRP)结合荧光标记抗体与检测探针上的荧光素标记结合,并加入四甲基联苯胺(TMB)底物,可检测这个复合体。用安培计测量由HRP介导的电子转运,信号与靶体的量成正比。

通过形成大量液体移动和局部加热,交流(AC)电动学提高了生物传感器测定的整体信噪比。进一步实施电动学原理将便于把生物传感器集成进POC设备,因为它不需要外部恒温器进行杂交。为了检测血吸虫,科学家用函数发生器在工作电极和辅助电极之间施加方波交流电势。

研究人员总结说,他们致力于开发快速检测尿液中埃及血吸虫卵的POC设备,目前已取得重大进步。他们实施的策略将有助于设备集成,如机械裂解和交流电动学。在将来的开发中,他们将把这一核心测定集成进全自动微流控盒,进一步优化检测灵敏度,并用临床样本进行确证。

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