Enhancing Point-of-Care Diagnosis of African Swine Fever Virus (ASFV) DNA with the CRISPR-Cas12a-Assisted Triplex Amplified Assay
文献类型： 外文期刊
作者： Zhu, Dan 1 ; Su, Tong 1 ; Sun, Tao 1 ; Qin, Xingcai 1 ; Su, Shao 1 ; Bai, Yun 4 ; Li, Fang 3 ; Zhao, Dongming 3 ; Shao, Guoqing 4 ; Chao, Jie 1 ; Feng, Zhixin 4 ; Wang, Lianhui 1 ;
作者机构： 1.Nanjing Univ Posts & Telecommun, Inst Adv Mat IAM, State Key Lab Organ Elect & Informat Displays, Nanjing 210023, Peoples R China
2.Nanjing Univ Posts & Telecommun, Inst Adv Mat IAM, Jiangsu Key Lab Biosensors, Nanjing 210023, Peoples R China
3.Chinese Acad Agr Sci, Harbin Vet Res Inst, State Key Lab Anim Dis Control & Prevent, Natl High Containment Facil Anim Dis Control & Pre, Harbin 150069, Peoples R China
4.Jiangsu Acad Agr Sci, Inst Vet Med, Minist Agr, Key Lab Vet Biol Engn & Technol, Nanjing 210014, Peoples R China
期刊名称：ANALYTICAL CHEMISTRY （ 影响因子：7.4； 五年影响因子：7.0 ）
ISSN： 0003-2700
年卷期： 2024 年 96 卷 13 期
页码：
收录情况： SCI
摘要： Accurate, ultrasensitive, and point-of-care (POC) diagnosis of the African swine fever virus (ASFV) remains imperative to prevent its spread and limit the losses incurred. Herein, we propose a CRISPR-Cas12a-assisted triplex amplified colorimetric assay for ASFV DNA detection with ultrahigh sensitivity and specificity. The specific recognition of recombinase aided amplification (RAA)-amplified ASFV DNA could activate the Cas12a/crRNA/ASFV DNA complex, leading to the digestion of the linker DNA (bio-L1) on magnetic beads (MBs), thereby preventing its binding of gold nanoparticles (AuNPs) network. After magnetic separation, the release of AuNPs network comprising a substantial quantity of AuNPs could lead to a discernible alteration in color and significantly amplify the plasmonic signal, which could be read by spectrophotometers or smartphones. By combining the RAA, CRISPR/Cas12a-assisted cleavage, and AuNPs network-mediated colorimetric amplification together, the assay could detect as low as 0.1 copies/mu L ASFV DNA within 1 h. The assay showed an accuracy of 100% for the detection of ASFV DNA in 16 swine tissue fluid samples, demonstrating its potential for on-site diagnosis of ASFV.
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