There is a critical need for rapid, sensitive, cost-effective, and portable detection systems for on-site measurement of ochratoxin A (OTA). Aptamers hold significant potential for portable biosensors, as liquid crystal (LC)-based aptasensors require no expensive lab equipment. Biosensors utilizing LC technology offer an encouraging approach for the creation of portable measurement systems, thereby removing the necessity for intricate and expensive laboratory apparatus. Furthermore, CRISPR technology is an excellent choice for enhancing diagnostic methods due to its portability, high accuracy, and sensitivity. This study introduces a three-way DNA junction (TWJ) that induces spatial disruption in the vertical arrangement of LC molecules, acting as an effective signal amplifier. This CRISPR-mediated biosensor detects OTA toxin in a wide linear range of 4.9 pg mL− 1 to 20 ng mL− 1 (R2=0.991), with a great detection limit of 1.47 pg mL− 1. It has been effectively used for detecting real samples of coffee, serum, and red grape juice, demonstrating its broad applications in agriculture and the food industry. The conceptual basis of this detection technique enables the creation of diverse aptasensors tailored for specific uses through the replacement of suitable sequences.

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