Dual real-time fluorescence saltatory rolling circle amplification (SRCA) for the simultaneous detection of soybean and peanut allergens in food

Soybeans and peanuts are nutritionally dense and are frequently incorporated into food products to enhance their nutritional profile and improve overall quality. However, soybeans and peanuts are among the primary causes of food allergies, with the potential to cause organ failure and even shock in severe cases. In this study, we combined saltatory rolling circle amplification (SRCA) and real-time fluorescence (RF) technology to simultaneously detect soybean and peanut allergens using a melting curve method. The dual RF-SRCA method demonstrated a sensitivity of 20 fg/μL, and the detection limits (LOD) of wheat flour spiked samples was determined to be 0.1% using this method. Additionally, the dual RF-SRCA reaction was demonstrated to be an effective method for the detection of soybean and peanut allergens. The relative sensitivity, specificity and accuracy of this method were 100%, 81.8%, and 96.7%. Consequently, this dual RF-SRCA test offers an accurate, reliable, and cost-effective method for the simultaneous detection of multiple allergens in food.