基于早孕试纸平台检测人唾液中基质金属蛋白酶-1,Analytical Chemistry

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基于早孕试纸平台检测人唾液中基质金属蛋白酶-1
Analytical Chemistry ( IF 6.7 ) Pub Date : 2022-11-15 , DOI: 10.1021/acs.analchem.2c03633
Zi-Tao Zhong ₁ , Ghazala Ashraf ₁ , Wei Chen ₁ , Bo Liu ₁ , Guo-Ping Wang ₁ , Yuan-Di Zhao _{1,

2}

Affiliation

基质金属蛋白酶（MMP）与肿瘤的发生和发展密切相关。建立一种低成本、简单、快速、灵敏的检测方法是口腔癌广谱筛查的迫切需要。在此，我们首次将MMP特异性切割能力与磁分离技术和商业试纸相结合，构建了一种灵敏的生物传感器，可以方便地检测MMP-1。该方法涉及两个 DNA 探针，肽-DNA1 和 hCG-DNA2，其中 DNA1 和 DNA2 是互补序列，生物素标记的肽可以稳定地结合链霉亲和素修饰的磁性纳米粒子。人绒毛膜促性腺激素 (hCG) 是妊娠试纸的目标。由 MMP-1 介导的裂解反应将肽-DNA1 和杂交的 hCG-DNA2 释放到溶液中，溶液中的hCG探针磁选后可在测定MMP-1的试纸条上显色。该方法利用MMP-1蛋白水解酶切的高特异性和试纸对靶标探针的高灵敏度，实现了MMP-1的灵敏检测，目视检测限为65.5 pg/mL。该方法在生物样品基质的应用中表现出比酶联免疫吸附测定更好的抗干扰性和灵敏度，表明其在临床诊断，特别是广谱口腔癌筛查方面具有巨大潜力。实现了 MMP-1 的灵敏检测，视觉检测限为 65.5 pg/mL。该方法在生物样品基质的应用中表现出比酶联免疫吸附测定更好的抗干扰性和灵敏度，表明其在临床诊断，特别是广谱口腔癌筛查方面具有巨大潜力。实现了 MMP-1 的灵敏检测，视觉检测限为 65.5 pg/mL。该方法在生物样品基质的应用中表现出比酶联免疫吸附测定更好的抗干扰性和灵敏度，表明其在临床诊断，特别是广谱口腔癌筛查方面具有巨大潜力。

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Detection of Matrix Metalloproteinase-1 in Human Saliva Based on a Pregnancy Test Strip Platform

Matrix metalloproteinase (MMP) is closely correlated with tumorigenesis and progression. Establishing a low-cost, simple, rapid, and sensitive method for its detection is highly desired for the broad-spectrum screening of oral cancer. Herein, we combine the MMP-specific cleavage ability with magnetic separation technology and a commercial test strip to construct a sensitive biosensor to detect MMP-1 conveniently for the first time. The method involves two DNA probes, peptide–DNA1 and hCG–DNA2, where DNA1 and DNA2 are complementary sequences, and the peptide labeled with biotin can bind streptavidin-modified magnetic nanoparticles stably. The human chorionic gonadotropin (hCG) is the target of the pregnancy test strip. The cleavage reaction mediated by MMP-1 releases peptide–DNA1 and the hybridized hCG–DNA2 into the solution, and the hCG probe in the solution can develop color on the test strip for the determination of MMP-1 after magnetic separation. This method utilizes the high specificity of MMP-1’s proteolytic cleavage and the high sensitivity of the test strip to the target probe, achieving a sensitive detection of MMP-1 with a visual detection limit of 65.5 pg/mL. The method shows better anti-interference and sensitivity than the enzyme-linked immunosorbent assay in the application of a biological sample matrix, suggesting its great potential for clinical diagnosis, especially for broad-spectrum oral cancer screening.

更新日期：2022-11-15

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