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Label-free, PCR-free chip-based detection of telomerase activity in bladder cancer cells

机译:基于无标记,无PCR的芯片检测膀胱癌细胞中的端粒酶活性

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Bladder cancer is one of the most common cancers in Worldwide. The determination of urinary telomerase activity is a promising tool for the diagnosis of bladder carcinoma owing to the high rate of expression of telomerase in cancer cells. Typical assay for telomerase activity is the telomeric repeat amplification protocol (TRAP) based on polymerase chain reaction (PCR) amplification. However, TRAP assay is susceptible to PCR-derived artifacts and requires time-consuming procedure, expensive equipments and reagents. To develop a new method for telomerase activity assay that is fast, simple, and cost-effective, we have examined a silicon-based microring resonator biosensor to detect label-free, PCR-free telomerase activity using telomerase extracted from two bladder cancer cell lines, J-82 and HT-1376, in a buffer solution and spiked urine. With telomerase primer immobilized microring resonator sensor system, we successfully demonstrate the detection of telomerse extracted from as little as 10. cells/μL and 100. cells/μL in buffer and urine, respectively. Especially, the results represented here is the first demonstration of the detection of telomerase activity in human urine on the chip-based system. From the results, we expect that the silicon photonic microring resonator system can provide a powerful tool for cost-effective and sensitive telomerase activity detection in urinary bladder cancer.
机译:膀胱癌是全球最常见的癌症之一。尿端粒酶活性的测定是诊断膀胱癌的有前途的工具,因为癌细胞中端粒酶的表达率很高。端粒酶活性的典型检测方法是基于聚合酶链反应(PCR)扩增的端粒重复扩增协议(TRAP)。然而,TRAP测定法容易受到PCR衍生的假象的影响,并且需要耗时的过程,昂贵的设备和试剂。为了开发一种快速,简单且经济高效的端粒酶活性测定新方法,我们研究了一种基于硅的微环共振器生物传感器,该传感器使用从两个膀胱癌细胞系中提取的端粒酶来检测无标签,无PCR的端粒酶活性。 ,J-82和HT-1376,放在缓冲溶液和加标尿液中。使用固定有端粒酶引物的微环共振器传感器系统,我们成功地证明了在缓冲液和尿液中分别从低至10.细胞/μL和100.细胞/μL提取的端粒酶的检测。特别是,此处表示的结果是在基于芯片的系统上检测人尿中端粒酶活性的第一个证明。从结果来看,我们期望硅光子微环谐振器系统可以为膀胱癌的经济有效且灵敏的端粒酶活性检测提供强大的工具。

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