1.延安大学化学化工学院,陕西 延安 716000
2.延安大学医学院,陕西 延安 716000
李淼(1999年生),女;研究方向:发光分析;E-mail:18932826802@yau.edu.cn
张越诚(1990年生),男;研究方向:生化分析;E-mail:yuechengzhang@yau.edu.cn
收稿:2026-04-15,
修回:2026-05-20,
录用:2026-05-27,
网络首发:2026-07,
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李淼, 高振耀, 张雅蓉, 等. DNA酶级联放大比色传感器用于细胞Mn2+检测[J/OL]. 中山大学学报(自然科学版)(中英文), 2026,1-10. DOI: 10.11714/acta.snus.ZR20260095.
Li Miao, Gao Zhenyao, Zhang Yarong, et al. DNAzyme-based cascade amplification colorimetric sensor for cellular Mn²⁺ detection[J/OL]. Acta Scientiarum Naturalium Universitatis Sunyatseni, 2026, 1-10.
李淼, 高振耀, 张雅蓉, 等. DNA酶级联放大比色传感器用于细胞Mn2+检测[J/OL]. 中山大学学报(自然科学版)(中英文), 2026,1-10. DOI: 10.11714/acta.snus.ZR20260095. DOI:
Li Miao, Gao Zhenyao, Zhang Yarong, et al. DNAzyme-based cascade amplification colorimetric sensor for cellular Mn²⁺ detection[J/OL]. Acta Scientiarum Naturalium Universitatis Sunyatseni, 2026, 1-10. DOI: 10.11714/acta.snus.ZR20260095.
锰离子(Mn
2+
)稳态失衡与多种疾病密切相关,但现有检测方法依赖大型仪器、操作繁琐,难以满足快速、便捷的分析需求。本研究基于Mn
2+
特异性DNA酶(Mn5V)与G-四链体/氯化血红素的级联信号放大原理,构建了一种高灵敏、高选择性的比色生物传感
器,用于复杂生物基质中Mn
2+
的检测。该传感器采用“发夹锁闭”式底物探针,在Mn
2+
存在时,Mn5V被激活并特异性切割底物链,释放出的DNA片段折叠形成G-四链体,进而与氯化血红素结合形成具有过氧化物酶活性的复合物,催化TMB-H
2
O
2
显色反应,实现信号的可视化输出。实验结果表明,该传感器对Mn
2+
表现出优异的分析性能,在7.0×10
-8
~ 1.0×10
-5
mol/L范围内呈现良好线性关系,检出限低至1.69×10
-9
mol/L,且对常见共存金属离子具有较强的抗干扰能力。将该传感器应用于MCF-7和A549细胞裂解液中的加标回收实验,回收率为98.00%~101.00%,相对标准偏差小于4.3%,验证了其在复杂生物样本中的准确性与可靠性。本研究构建的级联信号放大比色传感器为环境监测及锰代谢异常相关疾病的机制研究提供了一种新型、高效的分析工具。
Dysregulation of manganese ion (Mn
2+
) homeostasis is closely associated with various diseases. However, the existing detection methods rely on expensive instruments and involve cumbersome operations, making it difficult to meet the demands for rapid and convenient analysis. In this study, a highly sensitive and selective colorimetric biosensor was constructed based on the cascade signal amplification principle of Mn
2+
-specific DNAzyme (Mn5V) and G-quadruplex/hemin DNAzyme for the detection of Mn
2+
in complex biological matrices. The sensor employs a "hairpin-locked" substrate probe design. In the presence of Mn
2+
, Mn5V is activated and specifically cleaves the substrate strand, releasing a DNA fragment that folds into a G-quadruplex structure. The G-quadruplex then binds with hemin to form a complex with peroxidase-like activity, which catalyzes the TMB-H
2
O
2
colorimetric reaction, enabling visual signal output. Our results demonstrated that the sensor exhibits excellent analytical performance for Mn
2+
, showing a good linear relationship in the range of 7.0×10
-8
to 1.0×10
-5
mol/L with a detection limit as low as 1.69×10
-9
mol/L. The sensor also showed strong anti-interference ability against common coexisting metal ions. When applied to spiked recovery experiments in MCF-7 and A549 cell lysates, the recovery rates ranged from 98.00% to 101.00% with relative standard deviatio
ns below 4.3%, verifying its accuracy and reliability in complex biological samples. The cascade signal amplification colorimetric sensor constructed in this study provides a novel and efficient analytical tool for environmental monitoring and mechanistic research on the diseases related with manganese metabolism disorders.
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