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南京邮电大学汪联辉&苏邵&朱丹‖【Biosens Bioelectron】CRISPR驱动的DNA四面体介导传感界面用于猴痘病毒超灵敏检测

  • 2026-06-21 05:15:14
南京邮电大学汪联辉&苏邵&朱丹‖【Biosens Bioelectron】CRISPR驱动的DNA四面体介导传感界面用于猴痘病毒超灵敏检测

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介绍 /Introduction

猴痘病毒（MPXV）的快速、超灵敏检测对公共卫生安全至关重要。尽管聚合酶链反应（PCR）是当前诊断的金标准，但其依赖实验室基础设施、专业人员和昂贵设备，难以在资源匮乏地区推广。电化学生物传感器因其操作简便、成本低、灵敏度高、易于小型化和设备集成等优势，成为理想的即时检测（POCT）平台。近年来，CRISPR/Cas系统被广泛用于核酸检测，但多数CRISPR生物传感器仍需预扩增步骤（如PCR、LAMP等），存在污染风险、检测时间延长和设备依赖等问题。
为此，南京邮电大学汪联辉教授、苏邵教授和朱丹教授团队设计了一种无需预扩增的CRISPR驱动电化学生物传感器。该传感器整合了CRISPR/Cas12a系统、四面体DNA纳米结构（TDN）介导的传感界面和杂交链式反应（HCR）信号放大策略。TDN的三维刚性结构显著提高了探针的可及性和CRISPR的切割效率；靶标MPXV DNA激活Cas12a的反式切割活性，切断TDN上的初始链，从而阻止HCR反应的发生；反之，无靶标时HCR生成超长双链DNA，负载大量辣根过氧化物酶（HRP），催化TMB-H₂O₂体系产生显著放大的电化学信号。结合便携式电化学设备和智能手机，该传感器实现了对MPXV DNA的超灵敏、实时、现场检测，检测限低至194.48 aM，并在10%人唾液中表现出优异的回收率。这项工作为传染病的早期诊断提供了一种通用且强大的POCT平台。
相关成果以 “ Ultrasensitive detection of monkeypox virus: harnessing synergistic CRISPR-driven signal amplification on a DNA tetrahedron-mediated sensing interface” 为题发表在《Biosensors and Bioelectronics》。
「全文速递」南京邮电大学汪联辉&苏邵&朱丹‖BiosensBioelectronCRISPR驱动的DNA四面体介导传感界面用于猴痘病毒超灵敏检测.pdf

研究图文/Introduction

Scheme 1. Schematic illustration of a CRISPR-driven electrochemical biosensor for POCT detection of MPXV.

Fig. 1. (A) Polyacrylamide gel electrophoresis (PAGE) analysis of DNA tetrahedron (TDN) assembly. (B) PAGE analysis of the TDN-triggered hybridization chain reaction (HCR). (C) PAGE and (D) fluorescence spectra demonstrated the trans-cleavage activity of the CRISPR-Cas12a system. (E) Cyclic voltammetry (CV) curves of (a) SPCE-HFGNs, (b) TDN/SPCE-HFGNs, (c) MCH/TDN/SPCE-HFGNs, (d) BSA/MCH/TDN/SPCE-HFGNs, (e) H1+H2/BSA/MCH/TDN/SPCE-HFGNs, and (f) H1+H2/BSA/MCH/Cas12a-crRNA-MPXV DNA/TDN/SPCE-HFGNs in 5 mM [Fe(CN)6]3-/4- solution containing 0.1 M KCl. Scan rate: 100 mV s- 1. (F) Chronoamperometric responses of the biosensor with and without MPXV DNA.

Fig. 2. (A) Schematic illustration of three CRISPR-driven sensing strategies for MPXV DNA detection (i) TDN-HCR, (ii) ssDNA-HCR, and (iii) TDN-H1. Chronoamperometric responses of three CRISPR-driven sensing strategies (B) TDN-HCR, (C) ssDNA-HCR, and (D) TDN-H1 for detecting different concentrations of MPXV DNA. Calibration curves of (E) TDN-HCR, (F) ssDNA-HCR and (G) TDN-H1 for different concentrations of MPXV DNA detection. (H) ΔI of three CRISPR-driven sensing strategies for 100 pM MPXV DNA detection. (I) The corresponding LODs obtained from three sensing strategies. (J) Kinetic curves of TDN and ssDNA sensing interface for the trigger of HCR.

Fig. 3. The (A) selectivity, (B) reproducibility, and (C) stability of the CRISPR-driven electrochemical biosensor for MPXV DNA detection.
Fig. 4. (A) A smartphone-based portable electrochemical device for MPXV DNA detection. (B) Chronoamperometric curves of the portable device for detecting varying concentrations of MPXV DNA. (C) The linear relationship between the peak current and the logarithm of MPXV DNA concentration. (D) The performance of the portable device for MPXV DNA detection in 10 % human saliva samples.

结论 /Conclusion

该传感器整合了CRISPR/Cas12a系统、四面体DNA纳米结构传感界面和杂交链式反应信号放大策略。得益于SPCE-HFGNs（分级花状金纳米结构修饰的丝网印刷碳电极）的高导电性和比表面积、TDN的三维定向调控能力以及HCR的无酶信号放大效应，该传感器展现出宽动态范围（1 fM – 100 pM）、超低检测限（194.48 aM）、快速检测速度（30分钟内达到电流平台）、高重现性（RSD=3.59%）和优异的储存稳定性（7天后保持93%初始信号）。更重要的是，通过与便携式电化学设备和智能手机结合，该传感器成功实现了在10%人唾液中的加标回收实验，回收率在98.7%–108.8%之间，RSD小于5%，证明了其在实际样本中的POCT应用潜力。总体而言，这项工作为猴痘病毒的超灵敏现场检测提供了一个多功能且强大的平台，对传染病的早期诊断具有广阔的前景。

文献直通车 /Literature information

Ultrasensitive detection of monkeypox virus: harnessing synergistic CRISPR-driven signal amplification on a DNA tetrahedron-mediated sensing interface

Tianchen Du, Feng Ding, Xiaoxin Ma, Yi Luo, Dan Zhu,* Lianhui Wang,* Shao Su* Biosensors and Bioelectronics, Volume 298, 2026, 118441

原文链接：https://doi.org/10.1016/j.bios.2026.118441

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  1. CONNECT:[ UseTime:0.000457s ] mysql:host=127.0.0.1;port=3306;dbname=b460;charset=utf8mb4
  2. SHOW FULL COLUMNS FROM `fenlei` [ RunTime:0.000666s ]
  3. SELECT * FROM `fenlei` WHERE `fid` = 0 [ RunTime:0.004334s ]
  4. SELECT * FROM `fenlei` WHERE `fid` = 63 [ RunTime:0.000313s ]
  5. SHOW FULL COLUMNS FROM `set` [ RunTime:0.000766s ]
  6. SELECT * FROM `set` [ RunTime:0.000247s ]
  7. SHOW FULL COLUMNS FROM `article` [ RunTime:0.000623s ]
  8. SELECT * FROM `article` WHERE `id` = 581578 LIMIT 1 [ RunTime:0.009375s ]
  9. UPDATE `article` SET `lasttime` = 1783324979 WHERE `id` = 581578 [ RunTime:0.018604s ]
  10. SELECT * FROM `fenlei` WHERE `id` = 65 LIMIT 1 [ RunTime:0.000292s ]
  11. SELECT * FROM `article` WHERE `id` < 581578 ORDER BY `id` DESC LIMIT 1 [ RunTime:0.001961s ]
  12. SELECT * FROM `article` WHERE `id` > 581578 ORDER BY `id` ASC LIMIT 1 [ RunTime:0.024022s ]
  13. SELECT * FROM `article` WHERE `id` < 581578 ORDER BY `id` DESC LIMIT 10 [ RunTime:0.027005s ]
  14. SELECT * FROM `article` WHERE `id` < 581578 ORDER BY `id` DESC LIMIT 10,10 [ RunTime:0.051344s ]
  15. SELECT * FROM `article` WHERE `id` < 581578 ORDER BY `id` DESC LIMIT 20,10 [ RunTime:0.037394s ]
0.258380s