An Inexpensive CRISPR-Based Point-of-Care Test for the Identification of Meat Species and Meat Products
Dagang Tao, Xiao Xiao, Xiaochen Lan, Bingrong Xu, Yuan Wang, Emmanuel Mulaya Khazalwa, Wenya Pan, Jinxue Ruan, Yu Jiang, Xiangdong Liu, Changchun Li, Ruizhen Ye, Xinyun Li, Jing Xu, Shuhong Zhao, Shengsong Xie
Abstract
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.