An inexpensive CRISPR-based point-of-care test for the identification of meat species and meat products

The growing demand for and supply of meat and meat products has led to a proportional increase in cases of meat adulteration. Adulterated meat poses serious economic and health consequences globally. Current laboratory methods for meat species identification require specialized equipment with limite...

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Main Authors: Tao, D., Xiao, X., Lan, X., Xu, B., Wang, Y., Khazalwa, Emmanuel M., Pan, W., Ruan, J., Jiang, Y., Liu, X., Li, C., Ye, R., Li, X., Xu, J., Zhao, S., Xie, S.
Format: Journal Article
Language:Inglés
Published: MDPI 2022
Subjects:
Online Access:https://hdl.handle.net/10568/119686
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author Tao, D.
Xiao, X.
Lan, X.
Xu, B.
Wang, Y.
Khazalwa, Emmanuel M.
Pan, W.
Ruan, J.
Jiang, Y.
Liu, X.
Li, C.
Ye, R.
Li, X.
Xu, J.
Zhao, S.
Xie, S.
author_browse Jiang, Y.
Khazalwa, Emmanuel M.
Lan, X.
Li, C.
Li, X.
Liu, X.
Pan, W.
Ruan, J.
Tao, D.
Wang, Y.
Xiao, X.
Xie, S.
Xu, B.
Xu, J.
Ye, R.
Zhao, S.
author_facet Tao, D.
Xiao, X.
Lan, X.
Xu, B.
Wang, Y.
Khazalwa, Emmanuel M.
Pan, W.
Ruan, J.
Jiang, Y.
Liu, X.
Li, C.
Ye, R.
Li, X.
Xu, J.
Zhao, S.
Xie, S.
author_sort Tao, D.
collection Repository of Agricultural Research Outputs (CGSpace)
description The growing demand for and supply of meat and meat products has led to a proportional increase in cases of meat adulteration. Adulterated meat poses serious economic and health consequences globally. Current laboratory methods for meat species identification require specialized equipment with limited field applications. This study developed an inexpensive, point-of-care Loop-Mediated Isothermal Amplification (LAMP)-CRISPR/Cas12a colorimetric assay to detect meat species using a Texas Red-labelled single-strand (ssDNA) reporter. As low as 1.0 pg/µL of the porcine NADH4, the chicken NADH dehydrogenase subunit 2 (ND2) and the duck D-loop genes was detectable under white, blue and ultraviolet light. The test turnaround time from DNA extraction to visualization was approximately 40 min. The assay accurately detected pure and mixed-meat products in the laboratory (n = 15) and during a pilot point-of-care test (n = 8) in a food processing factory. The results are 100% reproducible using lateral flow detection strips and the real-time PCR detection instrument. This technology is fully deployable and usable in any standard room. Thus, our study demonstrates that this method is a straightforward, specific, sensitive, point-of-care test (POCT) adaptable to various outlets such as customs, quarantine units and meat import/export departments.
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spelling CGSpace1196862025-12-08T10:29:22Z An inexpensive CRISPR-based point-of-care test for the identification of meat species and meat products Tao, D. Xiao, X. Lan, X. Xu, B. Wang, Y. Khazalwa, Emmanuel M. Pan, W. Ruan, J. Jiang, Y. Liu, X. Li, C. Ye, R. Li, X. Xu, J. Zhao, S. Xie, S. animal products meat crispr The growing demand for and supply of meat and meat products has led to a proportional increase in cases of meat adulteration. Adulterated meat poses serious economic and health consequences globally. Current laboratory methods for meat species identification require specialized equipment with limited field applications. This study developed an inexpensive, point-of-care Loop-Mediated Isothermal Amplification (LAMP)-CRISPR/Cas12a colorimetric assay to detect meat species using a Texas Red-labelled single-strand (ssDNA) reporter. As low as 1.0 pg/µL of the porcine NADH4, the chicken NADH dehydrogenase subunit 2 (ND2) and the duck D-loop genes was detectable under white, blue and ultraviolet light. The test turnaround time from DNA extraction to visualization was approximately 40 min. The assay accurately detected pure and mixed-meat products in the laboratory (n = 15) and during a pilot point-of-care test (n = 8) in a food processing factory. The results are 100% reproducible using lateral flow detection strips and the real-time PCR detection instrument. This technology is fully deployable and usable in any standard room. Thus, our study demonstrates that this method is a straightforward, specific, sensitive, point-of-care test (POCT) adaptable to various outlets such as customs, quarantine units and meat import/export departments. 2022-05-19 2022-05-30T11:35:59Z 2022-05-30T11:35:59Z Journal Article https://hdl.handle.net/10568/119686 en Open Access MDPI Tao, D., Xiao, X., Lan, X., Xu, B., Wang, Y., Khazalwa, E.M., Pan, W., Ruan, J., Jiang, Y., Liu, X., Li, C., Ye, R., Li, X., Xu, J., Zhao, S. and Xie, S. 2022. An inexpensive CRISPR-based point-of-care test for the identification of meat species and meat products. Genes 13(5): 912.
spellingShingle animal products
meat
crispr
Tao, D.
Xiao, X.
Lan, X.
Xu, B.
Wang, Y.
Khazalwa, Emmanuel M.
Pan, W.
Ruan, J.
Jiang, Y.
Liu, X.
Li, C.
Ye, R.
Li, X.
Xu, J.
Zhao, S.
Xie, S.
An inexpensive CRISPR-based point-of-care test for the identification of meat species and meat products
title An inexpensive CRISPR-based point-of-care test for the identification of meat species and meat products
title_full An inexpensive CRISPR-based point-of-care test for the identification of meat species and meat products
title_fullStr An inexpensive CRISPR-based point-of-care test for the identification of meat species and meat products
title_full_unstemmed An inexpensive CRISPR-based point-of-care test for the identification of meat species and meat products
title_short An inexpensive CRISPR-based point-of-care test for the identification of meat species and meat products
title_sort inexpensive crispr based point of care test for the identification of meat species and meat products
topic animal products
meat
crispr
url https://hdl.handle.net/10568/119686
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