基于恒温指数扩增反应高灵敏度检测端粒酶活性
首发时间:2015-12-10
摘要:端粒酶延伸产物是具有(ggttag)n重复序列的DNA。设计一条与两个重复序列相匹配的特异性的DNA探针与端粒酶延伸产物杂交,形成双链DNA。过量的DNA探针被磁性石墨烯吸附,通过磁分离,被去除。利用恒温指数扩增反应(IEXPAR)对端粒酶延伸产物捕捉的DNA探针进行快速扩增。用与双链DNA特异性结合的SYBR Green I染料对扩增产物进行实时荧光检测。在优化条件下,可以检测到低至50个癌细胞的端粒酶活性。实现了恒温扩增条件下,端粒酶活性的高灵敏度检测。
关键词: 端粒酶活性 恒温指数扩增反应 磁性石墨烯 实时荧光检测
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Sensitive detection of telomerase activity based on isothermal exponetial amplification reaction
Abstract:Telomerase elongated product has many repeat sequences-(ggttag)n. A specific DNA probe which is matched with two repeat sequences is used to hybridize with telomerase elongated product. The excess DNA probes are attached to magnetic graphene and removed by magnetic separation. The DNA probes hybridized with telomerase elongated product is rapidly amplified by isothermal exponential amplification reaction (IEXPAR). SYBR Green I is utilized for real-time fluorescent detection of IEXPAR products. Under the optimal conditions, The telomerase activity in 50 cancer cells has been obviously detected. Sensitive detection of telomerase activity under the condition of constant temperature is achieved.
Keywords: telomerase activity isothermal exponential amplification reaction magnetic graphene real-time fluorescence detection
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