多色比色法在生物传感平台的研究进展

doi:10.19894/j.issn.1000-0518.230085

摘要/Abstract

摘要：

比色法是生物传感中较为常用的一种检测手段，因其具有较高的灵敏度和特异性而受到广泛关注。其中，基于多色探针的多色比色法已成为近年来的研究热点，灵敏度较单色比色法显著提高、且具有更高的分辨率和肉眼辨识度，通过观察颜色变化可进行可视化半定量检测，又可通过计算颜色比率或基于智能手机图像处理软件进行快速定量检测。基于有机小分子荧光显色、纳米材料显色及酶促反应显色3种不同的显色机理，综述了多色荧光小分子探针、多色纳米材料比色探针及多色酶促反应比色探针的检测机理以及它们在实际样本检测中的应用，并进一步探讨了其面临的挑战及未来发展机遇。

关键词: 荧光分子探针, 金纳米粒子, 银纳米粒子, 量子点, 酶联免疫吸附法, 纳米酶

Abstract:

The colorimetric method is a common detection method in biosensing， which has received much attention because of its high sensitivity and specificity. Among them， multicolor colorimetric method based on multicolor probes has become a hot research topic in recent years. Its sensitivity is significantly higher than that of monochromatic colorimetric method， and it has higher resolution which can be visualized for semi-quantitative detection by naked eye recognition， quickly quantified by calculating color ratios or based on smartphone image processing software. Based on three different color development mechanisms： organic small molecule fluorescence， nanomaterial color development and enzymatic reaction color development， the detection mechanisms of multicolor fluorescent small molecule probes， multicolor nanomaterial colorimetric probes and multicolor enzymatic reaction colorimetric probes are reviewed and their applications in practical sample detection are discussed. The challenges and future development opportunities are also discussed.

Key words: Fluorescent molecular probes, Gold nanoparticles, Silver nanoparticles, Quantum dots, Enzyme-linked immunosorbent assay, Nanoenzymes

中图分类号:

O652.7

徐一, 林梦瑶, 宫晓群. 多色比色法在生物传感平台的研究进展[J]. 应用化学, 2024, 41(1): 3-20.

Yi XU, Meng-Yao LIN, Xiao-Qun GONG. Research Progress of Multicolor Colorimetric Method in Biosensing Platform[J]. Chinese Journal of Applied Chemistry, 2024, 41(1): 3-20.

图/表 10

图1 双光子荧光分子探针原理及应用（A） Schematic diagram of the two-photon excitation principle［21］；（B） Schematic diagram of the principle of a flavonol-borate multicolor probe based on the detection of H2O2［26］；（C） Schematic diagram of the CQ-1 dual inspection probe［27］；（D） Schematic diagram of the RPC dual-detection probe principle［28］

Fig.1 Principles and applications of two-photon fluorescent probes

图2 多响应靶向型荧光分子探针应用（A， B） Multicolor fluorescent probes for targeting cellular lipid droplets［29］；（C） Multicolor fluorescence microimaging of NLS-LSS-mKate1 labelled nuclei （red） and GalT-ECFP labelled Golgi apparatus （blue） in tumour cells［30］；（D） Principle of the Mito-LX double detection probe［31］

Fig.2 Applications of multi-response targeted fluorescent probes

图3 多响应荧光寿命探针应用（A） FRET-FLIM imaging analysis reveals JKD-SCR interactions in endodermal cells［33］；（B） FLIM imaging compares the different cellular uptake mechanisms of OG-PTX［34］；（C） The NADH fluorescence lifetime of 4T1 cells in 2D culture （a） is shorter than that of 3D culture （b）， the black and red lines in （c） respectively represent the fluorescence lifetimes of NADH in 2D and 3D culture［36］

Fig.3 Applications of multi-response fluorescent lifetime probes

图4 聚集显色多色比色平台应用（A） Aggregation color changes of three functionalized AuNPs induced by Cys， GSH and GSSG and the corresponding UV-Vis spectra［41］；（B） Two-dimensional score plot of a categorised mercaptan multicolor colorimetric platform， demonstrating the platform's good discrimination of various mercaptans［41］

Fig.4 Applications of aggregated chromogenic multicolor platforms

图5 表面生长蚀刻多色比色平台应用（A） Schematic diagram and color change of AuNBPs growth method for detection of H5N1 virus［49］；（B） Schematic diagram and color change of AuNRs growth method for the detection of EA and vitamin C［50］；（C） Color change and concentration-absorbance change curves of AuNRs etching for cyanide detection［55］；（D） Schematic diagram and color change of AuNBPs etching method for the detection of telomerase activity［56］；（E） Schematic diagram and color change of AuNBP@MnO2 NSs etching method for amplification detection of exosomes［57］

Fig.5 Applications of multi-rcolor colorimetric platform for surface growth etching

图6 QDs多色比色平台应用（A） QD immunofluorescence imaging images and fluorescence spectra for cryoplethy smography of hepatocytes［63］；（B） Simultaneous detection of four toxins in individual and mixed fluorescence spectra by four-color QD［65］；（C） Schematic diagram of lateral flow strip for simultaneous detection of four nitrofuran metabolites by four-color QD［66］；（D） Schematic diagram of the principle of portable colorimetric detection of Hb by multicolor SiQDs［67］

Fig.6 Applications of multicolor colorimetric platform based on the QDs

图7 UCNP和MOF多色比色平台应用（A） Multicolor UCNP in vivo orthogonal imaging images［70］；（B） Schematic diagram of the two-color UCNP-LFT and portable reader for ST1 and BNP detection［74］；（C） Schematic diagram of the multi-color CDs-MOF sensing platform for the detection of Ag+ and Cys［75］；（D） Schematic diagram of a multi-color MOF sensing platform for the simultaneous detection of three types of DNA［76］

Fig.7 Applications of multicolor colorimetric platforms based on the UCNP and MOF

图8 蛋白酶催化的多色比色平台应用（A） Schematic diagram of the principle and color change of the multi-color ELISA for the detection of CBD［79］；（B） Color change of Listeria monocytogenes protease detected by multicolor lateral flow strip［81］?；（C） Schematic diagram of the principle and color change of a multicolor colorimetric platform for the detection of glucose by enzyme cascade amplification［83］?；（D） Schematic diagram of the principle and color change of the multicolor colorimetric platform for the detection of DDVP by enzyme cascade amplification［84］

Fig.8 Applications of protease-catalyzed multicolor colorimetric platforms

图9 基于DNA酶模拟物的多色比色平台应用（A） Schematic diagram of the principle of DNAzyme probes to detect miRNA in cells［97］；（B） Schematic diagram of the principle of multicolor detection of intracellular miRNA-21 and miRNA-155 by DNA orthotetrahedral probes［99］；（C） Schematic diagram of the principle of multicolor detection of PSA by DNA tridentate probes and film color change［100］

Fig.9 Applications of a multi-color colorimetric platform based on DNA enzyme simulants

图10 基于纳米酶的多色比色平台应用（A，B） Schematic diagram of the principle of VO2-QD naked eye multicolor detection of H2O2 and color change［104］；（C） Schematic diagram of the principle of multicolor fluorescence detection of ALP by Fef NCs［105］；（D） Schematic diagram of the principle of multicolor fluorescence detection of GSH by MnO2NS［106］；（E） Color changes in colorimetric cards for Pi， OPD and BA detected by MOF mimetic enzyme probes［107］；（F） Schematic diagram of the principle of SAN multicolor colorimetric strips for the detection of cardiac troponin I and myoglobin［108］

Fig.10 Applications of a nano-enzyme based multi-color colorimetric platform

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