CAJ | 학술논문

目的：建立一种基于纳米粒子的时间分辨（TR）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、高特异性地检测目标分子，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荧光传感器的检测限具有可比性。结论：基于纳米粒子的TR-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。
목적：건립일충기우납미입자적시간분변（TR）형광공진능량전이（FRET）생물전감기용우miRNAs적무확증검측。방법：이과핵감산단련probe I우련GdF3：Tb3+납미입자작위공체，이과핵감산단련probe II우련금（Au）납미입자작위수체，이용공체여수체지간적FRET검측목표miRNA hsa-miR-122-5p농도，병통과설치다공능매표의적검측연지거제자발형광배경이제고령민도。결과：해전감기능구고령민도、고특이성지검측목표분자，병차대우광조유량호적내수성，이차능구피면자발형광간우。대hsa-miR-122-5p적검측선성범위위0.1 fmol/L～100 pmol/L，여동류형적RNA형광전감기적검측한구유가비성。결론：기우납미입자적TR-FRET생물전감기용우miRNAs적무확증검측구유량호적령민도화특이성。
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 probeⅡas 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 Tb 3+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 type of sensors. 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.