A CRISPR/Cas12a-SERS platform for amplification-free detection of African swine fever virus genes

Huimin Wang; Ailing Su; Chengxin Bao; Chongyang Liang; Weiqing Xu; Jingjing Chang; Shuping Xu

doi:10.1016/j.talanta.2023.125225

A CRISPR/Cas12a-SERS platform for amplification-free detection of African swine fever virus genes

Talanta. 2024 Jan 15:267:125225. doi: 10.1016/j.talanta.2023.125225. Epub 2023 Sep 19.

Authors

Huimin Wang¹, Ailing Su², Chengxin Bao³, Chongyang Liang³, Weiqing Xu⁴, Jingjing Chang⁵, Shuping Xu⁶

Affiliations

¹ School of Chemistry and Environmental Engineering, Changchun University of Science and Technology, Changchun, 130022, PR China; State Key Laboratory of Supramolecular Structure and Materials, College of Chemistry, Jilin University, Changchun, 130012, PR China.
² State Key Laboratory of Supramolecular Structure and Materials, College of Chemistry, Jilin University, Changchun, 130012, PR China.
³ Institute of Frontier Medical Science, Jilin University, Changchun, 130021, PR China.
⁴ State Key Laboratory of Supramolecular Structure and Materials, College of Chemistry, Jilin University, Changchun, 130012, PR China; Theoretical Chemistry, College of Chemistry, Jilin University, Changchun, 130012, PR China.
⁵ School of Chemistry and Environmental Engineering, Changchun University of Science and Technology, Changchun, 130022, PR China. Electronic address: changjingjing@cust.edu.cn.
⁶ State Key Laboratory of Supramolecular Structure and Materials, College of Chemistry, Jilin University, Changchun, 130012, PR China; Theoretical Chemistry, College of Chemistry, Jilin University, Changchun, 130012, PR China; Center for Supramolecular Chemical Biology, College of Chemistry, Jilin University, Changchun, 130012, PR China. Electronic address: xusp@jlu.edu.cn.

PMID: 37741267
DOI: 10.1016/j.talanta.2023.125225

Abstract

A surface-enhanced Raman scattering (SERS) strategy combined with a CRISPR/Cas12a system is designed for the amplification-free gene detection of African swine fever virus (ASFV). A SERS sensing probe was fabricated by conjugating plasmonic SERS tags on the magnetic bead (MB) surface with an single-stranded DNA (ssDNA) as a linker. The target ASFV gene-activated Cas12a protein starts the trans-cleavage function on the linker ssDNA, which causes the release of SERS tags, leading to a decrease of the SERS signal detected above the collective MBs. Two signal enhancement strategies were adopted to improve the liquid-phase detection sensitivity arriving at the fM level. One is the unlimited trans-cleavage function of the Cas12a protein, and the other is the magnetic-induced collection of probes that can significantly gather the analytes from the solution to the laser spot and provide SERS hotspots during SERS measurement. Detection range is from 100 nM to 10 fM without the gene amplification steps. This sensing method achieved the SERS detection of ASFV gene in the serum system and the extracted nucleic acids in viral samples with high sensitivity and selectivity at a relative standard deviation of <8%. This sensing platform is mainly in use for site inspection and quick testing of gene samples.

Keywords: African swine fever virus; CRISPR/Cas12a; Gene detection; SERS.

MeSH terms

African Swine Fever Virus* / genetics
Animals
Biosensing Techniques*
CRISPR-Cas Systems
DNA, Single-Stranded
Nucleic Acids*
Swine

Substances

DNA, Single-Stranded
Nucleic Acids