糖化血红素
核酸外切酶
核酸外切酶 III
化学
检出限
血红蛋白
滚动圆复制
DNA–DNA杂交
荧光
生物系统
色谱法
分析化学(期刊)
生物物理学
DNA
分子生物学
生物化学
生物
物理
聚合酶
糖尿病
2型糖尿病
光学
大肠杆菌
基因
内分泌学
作者
Zhaopeng Chen,Ming-Li Su,Haoran Chen,Ruo Yuan,Yan Li,Wenbin Liang
标识
DOI:10.1016/j.bios.2022.113998
摘要
The glycated hemoglobin (HbA1c) level, defined as the ratio of HbA1c to total hemoglobin (tHb), has been considered as a standard index for diabetes mellitus diagnosis, avoiding any short-term fluctuation of the blood glucose level. However, it is still a key challenge that some complicated separation procedures are required to detect HbA1c and tHb in a single experiment. Herein, we developed a dual-sensing fluorescent assay for HbA1c and tHb based on proximity hybridization-induced rolling circle amplification (RCA) and T7 exonuclease aided signal cycle to calculate the HbAlc level in one single experiment. Typically, four DNA-tagged probes were used for immunoreactions of anti-HbA1c antibodies (Ab1) and anti-Hb antibodies (Ab2) with HbA1c and Hb, respectively. The RCA1 and RCA2 were induced by the complexes of assistant DNAs and the products of proximity hybridization with anti-HbA1c-DNA1 (2) and anti-Hb-DNA3 (4), respectively. Meanwhile, the tHb in solution could prevent the existing RCA2 based on its competing with Hb-DNA3 for Ab2-DNA4. Thus, RCA1 and RCA2 routes create "signal-on" and "signal-off" readouts, respectively. After the hybridization of signal probes and RCA products, the quenched fluorescence was recovered by the digestion of T7 exonuclease. Under optimized conditions, the method displayed high sensitivity with a limit of detection 2.17 ng/mL and 33.4 ng/mL for HbA1c and tHb, respectively, and the real sample analysis results were found to match well with the theoretical data, holding feasibility for rapid, homogeneous, and easy HbA1c level evaluation and great promise as a potential alternative tool to analyze HbA1c level clinically.
科研通智能强力驱动
Strongly Powered by AbleSci AI