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Silver nanoclusters-based fluorescent biosensing strategy for determination of mucin 1: combination of exonuclease I-assisted target recycling and graphene oxide-assisted hybridization chain reaction
Analytica Chimica Acta ( IF 5.7 ) Pub Date : 2020-09-01 , DOI: 10.1016/j.aca.2020.06.040
Hao Wu 1 , Jun Wu 1 , Yaling Liu 1 , Hongyong Wang 1 , Pei Zou 1
Affiliation  

A novel label-free fluorescent biosensing strategy was described for the sensitive detection of mucin 1 (MUC1). It consisted of an M-shaped aptamer probe for exonuclease I (Exo I)-assisted target recycling (EATR) amplification, and two AgNCs-hairpin probes for graphene oxide (GO)-assisted hybridization chain reaction (HCR) amplification. Based on the specificity of aptamer-target recognition, the addition of MUC1 caused a conformational change in the M-shaped aptamer probe, which was split into a MUC1-P3 complex and a P1-P2 duplex. Exo I then catalyzed the cleavage of aptamer sequence P3 from the MUC1-P3 complex and released the target MUC1. The released target MUC1 was free to bind with a new M-shaped probe to perform EATR amplification. Furthermore, the P1-P2 duplex with three single-stranded arms can act as a primer to initiate HCR between hairpin probes AgNCs-H1 and AgNCs-H2. In the process of HCR, two AgNCs-hairpins were autonomously cross-opened, generating long linear double-stranded nanowires containing large numbers of AgNCs. These nanowires cannot be quenched by GO due to the weak affinity between the long double-stranded DNA and GO, thereby retaining a strong fluorescent signal indicative of the concentration of MUC1. With these designs, in addition to an extremely low detection limit of 0.36 fg mL-1, the method exhibited an acceptable linear response to detect MUC1 from 1 fg mL-1 to 1 ng mL-1. Additionally, this method could be exerted with a high degree of success to detect MUC1 in diluted human serum with satisfactory results.

中文翻译:

基于银纳米团簇的荧光生物传感策略测定粘蛋白 1:结合外切核酸酶 I 辅助靶标回收和氧化石墨烯辅助杂交链反应

描述了一种新的无标记荧光生物传感策略,用于敏感检测粘蛋白 1 (MUC1)。它由一个用于外切核酸酶 I (Exo I) 辅助目标回收 (EATR) 扩增的 M 形适体探针和两个用于氧化石墨烯 (GO) 辅助杂交链反应 (HCR) 扩增的 AgNCs 发夹探针组成。基于适体-靶点识别的特异性,MUC1的加入引起了M形适体探针的构象变化,分裂为MUC1-P3复合体和P1-P2双链体。然后,Exo I 催化 MUC1-P3 复合物上的适体序列 P3 的裂解并释放目标 MUC1。释放的靶标 MUC1 自由结合新的 M 形探针进行 EATR 扩增。此外,具有三个单链臂的 P1-P2 双链体可以作为引物启动发夹探针 AgNCs-H1 和 AgNCs-H2 之间的 HCR。在 HCR 过程中,两个 AgNCs 发夹自动交叉打开,产生包含大量 AgNCs 的长线性双链纳米线。由于长双链 DNA 和 GO 之间的亲和力较弱,这些纳米线不能被 GO 淬灭,从而保留了指示 MUC1 浓度的强荧光信号。通过这些设计,除了 0.36 fg mL-1 的极低检测限外,该方法还表现出可接受的线性响应,可检测从 1 fg mL-1 到 1 ng mL-1 的 MUC1。此外,该方法可以高度成功地用于检测稀释人血清中的 MUC1,并获得令人满意的结果。两个 AgNCs 发夹自动交叉打开,产生包含大量 AgNCs 的长线性双链纳米线。由于长双链 DNA 和 GO 之间的亲和力较弱,这些纳米线不能被 GO 淬灭,从而保留了指示 MUC1 浓度的强荧光信号。通过这些设计,除了 0.36 fg mL-1 的极低检测限外,该方法还表现出可接受的线性响应,可检测 1 fg mL-1 至 1 ng mL-1 的 MUC1。此外,该方法可以高度成功地用于检测稀释人血清中的 MUC1,并获得令人满意的结果。两个 AgNCs 发夹自动交叉打开,产生包含大量 AgNCs 的长线性双链纳米线。由于长双链 DNA 和 GO 之间的亲和力较弱,这些纳米线不能被 GO 淬灭,从而保留了指示 MUC1 浓度的强荧光信号。通过这些设计,除了 0.36 fg mL-1 的极低检测限外,该方法还表现出可接受的线性响应,可检测从 1 fg mL-1 到 1 ng mL-1 的 MUC1。此外,该方法可以高度成功地用于检测稀释人血清中的 MUC1,并获得令人满意的结果。由于长双链 DNA 和 GO 之间的亲和力较弱,这些纳米线不能被 GO 淬灭,从而保留了指示 MUC1 浓度的强荧光信号。通过这些设计,除了 0.36 fg mL-1 的极低检测限外,该方法还表现出可接受的线性响应,可检测从 1 fg mL-1 到 1 ng mL-1 的 MUC1。此外,该方法可以高度成功地用于检测稀释人血清中的 MUC1,并获得令人满意的结果。由于长双链 DNA 和 GO 之间的亲和力较弱,这些纳米线不能被 GO 淬灭,从而保留了指示 MUC1 浓度的强荧光信号。通过这些设计,除了 0.36 fg mL-1 的极低检测限外,该方法还表现出可接受的线性响应,可检测 1 fg mL-1 至 1 ng mL-1 的 MUC1。此外,该方法可以高度成功地用于检测稀释人血清中的 MUC1,并获得令人满意的结果。
更新日期:2020-09-01
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