[1]张 毅,谭加兴,叶彦恺,等.基于纳米粒子的TR-FRET传感器用于miRNAs的无扩增检测[J].天津医科大学学报,2015,21(06):521-524.
ZHANG Yi,TAN Jia-xing,YE Yan-kai,et al. Direct non-amplification detection of miRNAs by time-resolved FRET sensor based on lanthanide-doped nanocrystals [J].Journal of Tianjin Medical University,2015,21(06):521-524.
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基于纳米粒子的TR-FRET传感器用于miRNAs的无扩增检测(PDF)
ZHANG Yi,TAN Jia-xing,YE Yan-kai,et al. Direct non-amplification detection of miRNAs by time-resolved FRET sensor based on lanthanide-doped nanocrystals [J].Journal of Tianjin Medical University,2015,21(06):521-524.
《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]
- 卷:
- 21卷
- 期数:
- 2015年06期
- 页码:
- 521-524
- 栏目:
- 技术与方法
- 出版日期:
- 2015-11-20
文章信息/Info
- 文章编号:
- 1006-8147（2015）06-0521-04
- Author(s):
- ZHANG Yi; TAN Jia-xing; YE Yan-kai; JIANG Wei; SUN Yan-hua; SHEN Wan-qiu; CAO Hai-yan; JIN Mei-na; QIN Nan; DUAN Hong-quan
- (Department of Medical Chemistry,?College of Pharmacy, Tianjin Medical University, Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and D（The然OSTIC说），Tianjin 300070, China)
- Keywords:
- lanthanide-doped' target="_blank" rel="external">">lanthanide-doped; nanocrystals; time-resolved; fluorescence resonance energy transfer; miRNA; sensor; non-amplification detection
- 分类号:
- R9
- DOI:
- -
- 文献标志码:
- A
- 摘要:
-
目的：建立一种基于纳米粒子的时间分辨荧光共振能量转移(FRET)生物传感器用于miRNAs的无扩增检测。方法：以寡核苷酸单链probe I 偶联GdF3:Tb3+纳米粒子作为供体，以寡核苷酸单链probe II 偶联金（Au）纳米粒子作为受体，利用供体与受体之间的FRET检测目标miRNA hsa-miR-122-5p浓度，并通过设置多功能酶标仪的检测延迟去除自发荧光背景以提高灵敏度。结果：该传感器能够高灵敏度、高特异性的检测目标分子，并且对于光照有良好的耐受性，而且能够避免自发荧光干扰。 对hsa-miR-122-5p的检测线性范围为0.1 fmol/L～100 pmol/L，与同类型的RNA荧光传感器的最佳检测限具有可比性。结论：基于纳米粒子的时间分辨FRET生物传感器用于miRNAs的无扩增检测具有良好的灵敏度和特异性。
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- Abstract:
- Objective: To develop a miRNA detection method based on the time-resolved (TR) fluorescence resonance energy transfer (FRET) assay. Methods: The GdF3:Tb3+ nanocrystals were coupled with nucleotide probe I as donor, and gold nanocrystals with nucleotide probe II as acceptor. miRNA hsa-miR-122-5p was detected by time-resolved FRET assay, and the short lived background luminescence such as auto-fluorescence was suppressed when determining the long lived fluorescence of Tb3+ by setting appropriate delay time and gate time. Results: The TR-FRET based miRNA sensor was sensitive and selective to hsa-miR-122-5p with a dynamic linear range of 0.1 fmol/L-100 pmol/L, which was compatible with that of the same of sensor. Furthermore, the GdF3:Tb3+ nanocrystals were stable to light irradiation and their lifetime was long enough to avoid autofluorescence. Conclusion: The TR-FRET miRNA sensor for non-amplification detection of hsa-miR-122-5p based on GdF3:Tb3+ nanocrystals, is sensitive and selective.
参考文献/References:
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备注/Memo
- 备注/Memo:
-
基金项目 国家自然科学基金资助项目（21305103，21373151，31200753，21205087）
作者简介 张毅（1982-），女，博士，研究方向：纳米材料于生化分析的应用；通信作者：段宏泉，E-mail: duanhq@tijmu.edu.cn。
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