基于CRISPR的即时检测平台用于鼻咽癌血浆EBV DNA筛查与监测
CRISPR-based point-of-care platform for EBV DNA screening and monitoring in nasopharyngeal cancer plasma.
文献信息
| PMID | 42826482 |
|---|---|
| 原文 | 在 PubMed 查看原文 ↗ |
| 发表日期 | 2026 |
| 作者 | Junheng Zhang |
| 作者单位 | Department of Laboratory Medicine, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, 200127, China. |
| 期刊 | Talanta |
| SCI 分区 | Q1 |
| IF | 6 |
| 研究类型 | AI/ML · 基础／转化 |
| 所属专科 | 鼻咽癌 |
中文摘要
背景： 血浆循环游离EB病毒（EBV）DNA是鼻咽癌（NPC）早期检测和纵向疾病监测的临床验证生物标志物。然而，当前的定量PCR（qPCR）检测需要复杂的仪器和专门的实验室基础设施，限制了其在资源有限环境中的可及性。方法：在此，我们提出了两种即时检测（POCT）平台，整合了RPA-CRISPR/Cas13a系统和油相磁珠转移（OMT）方法用于POCT检测：dμCRISPR，一种用于数字定量的液滴微流控芯片；以及idCRISPR，一种用于视觉筛查的集成侧向流装置。结果：OMT方法可在8分钟内从血浆中高效稳定地提取EBV DNA。dμCRISPR平台在60分钟内提供绝对定量结果，分析灵敏度为1拷贝/μL。在24份临床血浆样本的平行测试中，dμCRISPR与qPCR结果一致。此外，idCRISPR平台可在50分钟内实现裸眼筛查，检测限为10拷贝/μL，在评估的30份临床样本中显示与qPCR结果一致。结论：这些平台共同构成了精确定量和快速筛查的双轨策略，支持在不同医疗环境中对NPC的端到端管理。
英文摘要
Plasma circulating cell-free Epstein-Barr virus (EBV) DNA is a clinically validated biomarker for early nasopharyngeal carcinoma (NPC) detection and longitudinal disease monitoring. However, current quantitative PCR (qPCR) assays require sophisticated instrumentation and specialized laboratory infrastructure, constraining their accessibility in resource-limited settings. Here, we present two point-of-care testing platforms integrating the RPA-CRISPR/Cas13a system and oil-phase magnetic bead transfer (OMT) method for POCT detection: dμCRISPR, a droplet microfluidic chip for digital quantification, and idCRISPR, an integrated lateral flow device for visual screening. The OMT method achieves efficient and stable EBV DNA extraction from plasma within 8 min. The dμCRISPR platform provides absolute quantitative results within 60 min with an analytical sensitivity of 1 copy/μL. In parallel testing of 24 clinical plasma samples, dμCRISPR were consistent with qPCR. Additionally, the idCRISPR platform enables naked-eye screening within 50 min with a detection limit of 10 copies/μL, showing results consistent with qPCR in the 30 clinical samples evaluated. Together, these platforms form a dual-track strategy for precise quantification and rapid screening, supporting the end-to-end management of NPC across diverse healthcare environments.