Dual-amplification system based on CRISPR-Cas12a and horseradish peroxidase-tethered magnetic microspheres for colorimetric detection of microcystin-LR
Wu, Pian LU ; Zhang, Man LU ; Xue, Xiaoting LU ; Ding, Ping and Ye, Lei LU
(2023)
In Microchimica Acta
190(8).
- Abstract
A novel dual-amplification system based on CRISPR-Cas12a and horseradish peroxidase (HRP) was developed for colorimetric determination of MC-LR. This dual-amplification was accomplished by combining the nuclease activity of CRISPR-Cas12a with the redox activity of HRP. HRP linked to magnetic beads through an ssDNA (MB-ssDNA-HRP) was used to induce a color change of the 3,3′,5,5′-tetramethylbenzidine (TMB)-H2O2 chromogenic substrate solution. Specific binding of MC-LR with its aptamer initiated the release of a complementary DNA (cDNA), which was designed to activate the trans-cleavage activity of CRISPR-Cas12a. Upon activation, Cas12a cut the ssDNA linker in MB-ssDNA-HRP, causing a reduction of HRP on the magnetic... (More)
A novel dual-amplification system based on CRISPR-Cas12a and horseradish peroxidase (HRP) was developed for colorimetric determination of MC-LR. This dual-amplification was accomplished by combining the nuclease activity of CRISPR-Cas12a with the redox activity of HRP. HRP linked to magnetic beads through an ssDNA (MB-ssDNA-HRP) was used to induce a color change of the 3,3′,5,5′-tetramethylbenzidine (TMB)-H2O2 chromogenic substrate solution. Specific binding of MC-LR with its aptamer initiated the release of a complementary DNA (cDNA), which was designed to activate the trans-cleavage activity of CRISPR-Cas12a. Upon activation, Cas12a cut the ssDNA linker in MB-ssDNA-HRP, causing a reduction of HRP on the magnetic beads. Consequently, the UV–Vis absorbance of the HRP-catalyzed reaction was decreased. The dual-signal amplification facilitated by CRISPR-Cas12a and HRP enabled the colorimetric detection of MC-LR in the range 0.01 to 50 ng·mL−1 with a limit of detection (LOD) of 4.53 pg·mL−1. The practicability of the developed colorimetric method was demonstrated by detecting different levels of MC-LR in spiked real water samples. The recoveries ranged from 86.2 to 118.5% and the relative standard deviation (RSD) was 8.4 to 17.6%. This work provides new inspiration for the construction of effective signal amplification platforms and demonstrates a simple and user-friendly colorimetric method for determination of trace MC-LR.
(Less)
- author
- Wu, Pian
LU
; Zhang, Man
LU
; Xue, Xiaoting
LU
; Ding, Ping
and Ye, Lei
LU
- organization
- publishing date
- 2023
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Colorimetric detection, CRISPR-Cas12a, Horseradish peroxidase, Microcystin-LR
- in
- Microchimica Acta
- volume
- 190
- issue
- 8
- article number
- 314
- publisher
- Springer
- external identifiers
-
- pmid:37474872
- scopus:85165390748
- ISSN
- 0026-3672
- DOI
- 10.1007/s00604-023-05887-9
- language
- English
- LU publication?
- yes
- id
- 588a6543-4b50-4f56-9465-2e86c053ee00
- date added to LUP
- 2023-09-01 14:45:28
- date last changed
- 2025-10-20 13:55:37
@article{588a6543-4b50-4f56-9465-2e86c053ee00,
abstract = {{<p>A novel dual-amplification system based on CRISPR-Cas12a and horseradish peroxidase (HRP) was developed for colorimetric determination of MC-LR. This dual-amplification was accomplished by combining the nuclease activity of CRISPR-Cas12a with the redox activity of HRP. HRP linked to magnetic beads through an ssDNA (MB-ssDNA-HRP) was used to induce a color change of the 3,3′,5,5′-tetramethylbenzidine (TMB)-H<sub>2</sub>O<sub>2</sub> chromogenic substrate solution. Specific binding of MC-LR with its aptamer initiated the release of a complementary DNA (cDNA), which was designed to activate the trans-cleavage activity of CRISPR-Cas12a. Upon activation, Cas12a cut the ssDNA linker in MB-ssDNA-HRP, causing a reduction of HRP on the magnetic beads. Consequently, the UV–Vis absorbance of the HRP-catalyzed reaction was decreased. The dual-signal amplification facilitated by CRISPR-Cas12a and HRP enabled the colorimetric detection of MC-LR in the range 0.01 to 50 ng·mL<sup>−1</sup> with a limit of detection (LOD) of 4.53 pg·mL<sup>−1</sup>. The practicability of the developed colorimetric method was demonstrated by detecting different levels of MC-LR in spiked real water samples. The recoveries ranged from 86.2 to 118.5% and the relative standard deviation (RSD) was 8.4 to 17.6%. This work provides new inspiration for the construction of effective signal amplification platforms and demonstrates a simple and user-friendly colorimetric method for determination of trace MC-LR. <br/></p>}},
author = {{Wu, Pian and Zhang, Man and Xue, Xiaoting and Ding, Ping and Ye, Lei}},
issn = {{0026-3672}},
keywords = {{Colorimetric detection; CRISPR-Cas12a; Horseradish peroxidase; Microcystin-LR}},
language = {{eng}},
number = {{8}},
publisher = {{Springer}},
series = {{Microchimica Acta}},
title = {{Dual-amplification system based on CRISPR-Cas12a and horseradish peroxidase-tethered magnetic microspheres for colorimetric detection of microcystin-LR}},
url = {{http://dx.doi.org/10.1007/s00604-023-05887-9}},
doi = {{10.1007/s00604-023-05887-9}},
volume = {{190}},
year = {{2023}},
}