Hu Chao, Kong Xiang Juan, Yu Ru Qin, Chen Ting Ting, Chu Xia
State Key Laboratory of Chemo/Bio-Sensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University.
Anal Sci. 2017;33(7):783-788. doi: 10.2116/analsci.33.783.
A novel fluorescence sensing platform for ultrasensitive detection of S1 nuclease activity has been constructed based on MnO nanosheets and FAM labeled single-stranded DNA (FAM-ssDNA). In this system, MnO nanosheets were found to have different adsorbent ability toward ssDNA and mono- or oligonucleotide fragments. FAM-ssDNA could adsorb on MnO nanosheets and resulted in significant fluorescence quenching through fluorescence resonance energy transfer (FRET), while mono- or oligonucleotide fragments could not adsorb on MnO nanosheets and still retained strong fluorescence emission. With the addition of S1 nuclease, FAM-ssDNA was cleaved into mono- or oligonucleotide fragments, which were not able to adsorb on MnO nanosheets and the fluorescence signal was never quenched. The different fluorescence intensity allowed for examination of S1 nuclease activity. The developed method can detect S1 nuclease activity in the range of 0 - 20 U mL with a detection limit of 0.05 U mL. Benefits of the system include less time-consuming processes and more simple design compared to other endonuclease assays. Satisfactory performance for S1 nuclease in complex samples has been successfully demonstrated with the system. The developed assay could potentially provide a new platform in bioimaging and clinical diagnosis.
基于MnO纳米片和FAM标记的单链DNA(FAM-ssDNA)构建了一种用于超灵敏检测S1核酸酶活性的新型荧光传感平台。在该系统中,发现MnO纳米片对ssDNA以及单链或寡核苷酸片段具有不同的吸附能力。FAM-ssDNA可以吸附在MnO纳米片上,并通过荧光共振能量转移(FRET)导致显著的荧光猝灭,而单链或寡核苷酸片段不能吸附在MnO纳米片上,仍保留强烈的荧光发射。加入S1核酸酶后,FAM-ssDNA被切割成单链或寡核苷酸片段,这些片段不能吸附在MnO纳米片上,荧光信号也不会猝灭。荧光强度的差异可用于检测S1核酸酶的活性。所开发的方法能够在0-20 U/mL范围内检测S1核酸酶活性,检测限为0.05 U/mL。与其他内切核酸酶检测方法相比,该系统具有耗时更少、设计更简单的优点。该系统已成功证明在复杂样品中对S1核酸酶具有令人满意的检测性能。所开发的检测方法可能为生物成像和临床诊断提供一个新的平台。