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基于 DNA 框架信号放大平台的高通量系统免疫监测。

DNA framework signal amplification platform-based high-throughput systemic immune monitoring.

机构信息

State Key Laboratory of Oral Diseases, National Center for Stomatology, National Clinical Research Center for Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, 610041, Sichuan, PR China.

Department of Prosthodontics, Tianjin Stomatological Hospital, School of Medicine, Nankai University, Tianjin, 300041, PR China.

出版信息

Signal Transduct Target Ther. 2024 Feb 7;9(1):28. doi: 10.1038/s41392-024-01736-0.

Abstract

Systemic immune monitoring is a crucial clinical tool for disease early diagnosis, prognosis and treatment planning by quantitative analysis of immune cells. However, conventional immune monitoring using flow cytometry faces huge challenges in large-scale sample testing, especially in mass health screenings, because of time-consuming, technical-sensitive and high-cost features. However, the lack of high-performance detection platforms hinders the development of high-throughput immune monitoring technology. To address this bottleneck, we constructed a generally applicable DNA framework signal amplification platform (DSAP) based on post-systematic evolution of ligands by exponential enrichment and DNA tetrahedral framework-structured probe design to achieve high-sensitive detection for diverse immune cells, including CD4+, CD8+ T-lymphocytes, and monocytes (down to 1/100 μl). Based on this advanced detection platform, we present a novel high-throughput immune-cell phenotyping system, DSAP, achieving 30-min one-step immune-cell phenotyping without cell washing and subset analysis and showing comparable accuracy with flow cytometry while significantly reducing detection time and cost. As a proof-of-concept, DSAP demonstrates excellent diagnostic accuracy in immunodeficiency staging for 107 HIV patients (AUC > 0.97) within 30 min, which can be applied in HIV infection monitoring and screening. Therefore, we initially introduced promising DSAP to achieve high-throughput immune monitoring and open robust routes for point-of-care device development.

摘要

系统性免疫监测是通过定量分析免疫细胞来进行疾病早期诊断、预后和治疗规划的重要临床工具。然而,传统的使用流式细胞术的免疫监测在大规模样本测试中面临着巨大的挑战,特别是在大规模健康筛查中,因为它耗时、技术敏感且成本高。然而,缺乏高性能的检测平台阻碍了高通量免疫监测技术的发展。为了解决这一瓶颈问题,我们构建了一个普遍适用的 DNA 框架信号扩增平台 (DSAP),该平台基于配体的系统进化指数富集和 DNA 四面体型框架结构探针设计,实现了对多种免疫细胞(包括 CD4+、CD8+ T 淋巴细胞和单核细胞)的高灵敏度检测(低至 1/100 μl)。基于这个先进的检测平台,我们提出了一种新型的高通量免疫细胞表型分析系统 DSAP,实现了 30 分钟一步式免疫细胞表型分析,无需细胞洗涤和亚群分析,并且与流式细胞术相比具有相当的准确性,同时大大缩短了检测时间和成本。作为概念验证,DSAP 在 107 名 HIV 患者的免疫缺陷分期中表现出优异的诊断准确性(AUC>0.97),可在 30 分钟内完成,可用于 HIV 感染监测和筛查。因此,我们首次引入有前途的 DSAP 来实现高通量免疫监测,并为即时检测设备的开发开辟了广阔的途径。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/77d3/10847453/46ad8e9b17fe/41392_2024_1736_Fig1_HTML.jpg

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