Double Isothermal Amplification and CRISPR-Cas12a for Sensitive Detection of Citrinin
(2021) In ACS Agricultural Science and Technology 1(10). p.1997-2005- Abstract
- An analytical method is developed for ultrasensitive detection of citrinin using double isothermal amplification and CRISPR-Cas12a. Gold nanoparticles (AuNPs) modified with antigen and thiol-terminated, single-strand DNA (ssDNA) are used as a probe. The antigen-modified AuNPs compete with citrinin to bind to magnetic beads coated with an anticitrinin antibody. After a simple magnetic separation, the AuNPs are collected, and the ssDNA are released after they are washed with a dithiothreitol solution. The ssDNA is first amplified by an exponential amplification reaction and then used as a primer in a subsequent hybridization chain reaction to produce double-stranded DNA (dsDNA) that contains a protospacer adjacent motif to allow recognition... (More)
- An analytical method is developed for ultrasensitive detection of citrinin using double isothermal amplification and CRISPR-Cas12a. Gold nanoparticles (AuNPs) modified with antigen and thiol-terminated, single-strand DNA (ssDNA) are used as a probe. The antigen-modified AuNPs compete with citrinin to bind to magnetic beads coated with an anticitrinin antibody. After a simple magnetic separation, the AuNPs are collected, and the ssDNA are released after they are washed with a dithiothreitol solution. The ssDNA is first amplified by an exponential amplification reaction and then used as a primer in a subsequent hybridization chain reaction to produce double-stranded DNA (dsDNA) that contains a protospacer adjacent motif to allow recognition by CRISPR-Cas12a. The dsDNA activates the Cas12a-gRNA to cleave a reporter ssDNA to generate a fluorescence signal. The developed analytical method has a low detection limit (0.127 ng mL–1) and a wide linear range (0.005–500 μg mL–1) for detection of citrinin. For detection of citrinin in oat and flour, recoveries of 97–104% and 105–111% are obtained, respectively. By combining double isothermal amplification with CRISPR-Cas12a, ultrahigh sensitivity and selectivity can be achieved for detection of toxins in food. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/52ef5489-9d96-4a06-a214-8e841b2673ec
- author
- Zhang, Man LU ; Xue, Xiaoting LU ; Gong, Haiyue LU ; Liu, Baolin and Ye, Lei LU
- organization
- publishing date
- 2021-10-25
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Ultrasensitive detection, Biobarcode, CRISPR-Cas12a, Exponential amplification reaction, Hybridization chain reaction, Citrinin
- in
- ACS Agricultural Science and Technology
- volume
- 1
- issue
- 10
- pages
- 9 pages
- publisher
- The American Chemical Society (ACS)
- external identifiers
-
- scopus:85125573215
- ISSN
- 2692-1952
- DOI
- 10.1021/acsfoodscitech.1c00321
- language
- English
- LU publication?
- yes
- id
- 52ef5489-9d96-4a06-a214-8e841b2673ec
- date added to LUP
- 2022-09-29 09:26:28
- date last changed
- 2023-02-15 09:08:43
@article{52ef5489-9d96-4a06-a214-8e841b2673ec, abstract = {{An analytical method is developed for ultrasensitive detection of citrinin using double isothermal amplification and CRISPR-Cas12a. Gold nanoparticles (AuNPs) modified with antigen and thiol-terminated, single-strand DNA (ssDNA) are used as a probe. The antigen-modified AuNPs compete with citrinin to bind to magnetic beads coated with an anticitrinin antibody. After a simple magnetic separation, the AuNPs are collected, and the ssDNA are released after they are washed with a dithiothreitol solution. The ssDNA is first amplified by an exponential amplification reaction and then used as a primer in a subsequent hybridization chain reaction to produce double-stranded DNA (dsDNA) that contains a protospacer adjacent motif to allow recognition by CRISPR-Cas12a. The dsDNA activates the Cas12a-gRNA to cleave a reporter ssDNA to generate a fluorescence signal. The developed analytical method has a low detection limit (0.127 ng mL<sup>–1</sup>) and a wide linear range (0.005–500 μg mL<sup>–1</sup>) for detection of citrinin. For detection of citrinin in oat and flour, recoveries of 97–104% and 105–111% are obtained, respectively. By combining double isothermal amplification with CRISPR-Cas12a, ultrahigh sensitivity and selectivity can be achieved for detection of toxins in food.}}, author = {{Zhang, Man and Xue, Xiaoting and Gong, Haiyue and Liu, Baolin and Ye, Lei}}, issn = {{2692-1952}}, keywords = {{Ultrasensitive detection; Biobarcode; CRISPR-Cas12a; Exponential amplification reaction; Hybridization chain reaction; Citrinin}}, language = {{eng}}, month = {{10}}, number = {{10}}, pages = {{1997--2005}}, publisher = {{The American Chemical Society (ACS)}}, series = {{ACS Agricultural Science and Technology}}, title = {{Double Isothermal Amplification and CRISPR-Cas12a for Sensitive Detection of Citrinin}}, url = {{http://dx.doi.org/10.1021/acsfoodscitech.1c00321}}, doi = {{10.1021/acsfoodscitech.1c00321}}, volume = {{1}}, year = {{2021}}, }