CRISPR-Cas12-based field-deployable system for rapid detection of synthetic DNA sequence of the monkeypox virus genome

被引：46

作者：

Chen, Qun ^{[1
,2
]}

Gul, Ijaz ^{[1
,2
]}

Liu, Changyue ^{[1
,2
]}

Lei, Zhengyang ^{[1
,2
]}

Li, Xingyu

Raheem, Muhammad A. ^{[1
,2
,3
]}

He, Qian ^{[1
,2
]}

Zhang Haihui ^{[1
,2
]}

Leeansyah, Edwin ^{[1
,2
]}

Zhang, Can Y. ^{[1
,2
]}

Pandey, Vijay ^{[1
,2
]}

Du, Ke ^{[4
]}

Qin, Peiwu ^{[1
,2
]}

机构：

[1] Tsinghua Univ, Tsinghua Shenzhen Int Grad Sch, Inst Biopharmaceut & Hlth Engn, Shenzhen 518055, Peoples R China

[2] Tsinghua Univ, Tsinghua Berkeley Shenzhen Inst, Tsinghua Shenzhen Int Grad Sch, Shenzhen, Peoples R China

[3] Cent South Univ, Dept Hepatobiliary & Pancreat Surg 2, Xiangya Hosp 3, Changsha, Hunan, Peoples R China

[4] Univ Calif Riverside, Dept Chem & Environm Engn, Riverside, CA 92521 USA

来源：

JOURNAL OF MEDICAL VIROLOGY | 2023年 / 95卷 / 01期

基金：

中国国家自然科学基金;

关键词：

CRISPR-Cas12a; detection; monkeypox endemic; monkeypox virus; recombinase polymerase amplification; VARIOLA; ASSAY;

D O I：

10.1002/jmv.28385

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

The global outbreak of the monkeypox virus (MPXV) highlights the need for rapid and cost-effective MPXV detection tools to effectively monitor and control the monkeypox disease. Herein, we demonstrated a portable CRISPR-Cas-based system for naked-eye detection of MPXV. The system harnesses the high selectivity of CRISPR-Cas12 and the isothermal nucleic acid amplification potential of recombinase polymerase amplification. It can detect both the current circulating MPXV clade and the original clades. We reached a limit of detection (LoD) of 22.4aM (13.5copies/mu l) using a microtiter plate reader, while the visual LoD of the system is 75aM (45copies/mu l) in a two-step assay, which is further reduced to 25aM (15copies/mu l) in a one-pot system. We compared our results with quantitative polymerase chain reaction and obtained satisfactory consistency. For clinical application, we demonstrated a sensitive and precise visual detection method with attomolar sensitivity and a sample-to-answer time of 35min.

引用

页数：11

共 46 条

[31] Recombinase polymerase amplification: Basics, applications and recent advances [J].