Droplet digital polymerase chain reaction (ddPCR) for rapid screening of adulterants in honey: A case study on acacia honey adulterated with canola honey

Abstract

In this study, droplet digital polymerase chain reaction (ddPCR) was used to detect adulteration of acacia honey with canola honey. DNA extraction from pollen in acacia honey and canola honey was performed using four different pollen treatment methods, including, enzymatic hydrolysis, freeze-thaw, mechanical, and ultrasonic. The mechanical extraction method was the most preferred method of DNA extraction from honey. A duplex ddPCR method developed based on the specific target gene in acacia and canola was performed. The detection results showed that the established duplex ddPCR had a high detection sensitivity, detecting up to 1% adulteration. The accuracy of duplex ddPCR was consistent with the traditional melissopalynologic method. All results indicated that ddPCR was a powerful method for assessing the authenticity and quality of honey. • A pollen treatment method for extraction DNA from honey was proposed. • A duplex ddPCR based on the specific target gene in acacia and canola was developed. • The duplex ddPCR method had a high detection sensitivity, detecting up to 1% adulteration. • The method was applied to assess real samples to demonstrate the practicability for adulteration detection.