基于恒温指数扩增反应高灵敏度检测端粒酶活性

doi:10.3969/j.issn.1000-1565.2016.02.008

河北大学学报(自然科学版) ›› 2016, Vol. 36 ›› Issue (2): 155-161.DOI: 10.3969/j.issn.1000-1565.2016.02.008

基于恒温指数扩增反应高灵敏度检测端粒酶活性

周晓毓,张佳玉,周曼,贾红霞

收稿日期:2015-11-25 出版日期:2016-03-25 发布日期:2016-03-25
通讯作者: 贾红霞(1980—),女,河北宣化人,河北大学讲师,博士,主要从事生化分析及光谱分析研究.E-mail: jia123renren@126.com
作者简介:周晓毓(1990—),女,河北阜城人,河北大学在读研究生. E-mail: love_zhouxy@126.com
基金资助:
国家自然科学基金青年基金资助项目(21405032);教育部博士点基金资助项目(20121301120006)

Sensitive detection of telomerase activity based on isothermal exponetial amplification reaction

ZHOU Xiaoyu,ZHANG Jiayu,ZHOU Man,JIA Hongxia

College of Chemistry and Environmental Science, Hebei University, Baoding 071002, China

Received:2015-11-25 Online:2016-03-25 Published:2016-03-25

摘要/Abstract

摘要： 端粒酶延伸产物是具有(ggttag)_n重复序列的DNA.设计1条与2个重复序列相匹配的特异性的DNA探针-(ctaacc)₂使其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探针进行快速的扩增.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个癌细胞中的端粒酶活性.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

Abstract: Telomerase elongated product has many repeat sequences-(ggttag)_n.A specific DNA probe-(ctaacc)₂ which is matched with two repeat sequences is used to hybridize with telomerase elongated product.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 only 50 cancer cells can be easily detected.Sensitive detection of telomerase activity under isothermal condition is achieved.

Key words: telomerase activity, isothermal exponential amplification reaction, magnetic graphene, real-time fluorescence detection

中图分类号:

周晓毓,张佳玉,周曼,贾红霞. 基于恒温指数扩增反应高灵敏度检测端粒酶活性[J]. 河北大学学报(自然科学版), 2016, 36(2): 155-161.

ZHOU Xiaoyu,ZHANG Jiayu,ZHOU Man,JIA Hongxia. Sensitive detection of telomerase activity based on isothermal exponetial amplification reaction[J]. Journal of Hebei University (Natural Science Edition), 2016, 36(2): 155-161.

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基于恒温指数扩增反应高灵敏度检测端粒酶活性

Sensitive detection of telomerase activity based on isothermal exponetial amplification reaction

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