Abstract
Cashew nut allergy shows an increasing prevalence along with its high consumption. Nowadays, food allergy still has no cure, and allergen avoidance is still the standard management, urging a sensitive and convenient cashew nut detection method. Herein, saltatory rolling circle amplification (SRCA), loop-mediated isothermal amplification (LAMP), as well as polymerase chain reaction (PCR) targeting the Ana o 2 gene were proposed and compared. In addition, a closed-tube SRCA assay using hydroxynaphthol blue (HNB) as a color indicator (HNB-SRCA) was developed to realize visual on-site detection of cashew nut. SRCA and HNB-SRCA can detect 75.3 fg of cashew nut DNA, which is 10- and 100-times more sensitive than LAMP and PCR, respectively. SRCA and HNB-SRCA exhibited a relative sensitivity of 0.01% in a binary mixture, which is comparable to LAMP and 10-times higher than that of PCR. When combined with a rapid DNA extraction method, SRCA and HNB-SRCA performed better applicability compared to LAMP and PCR. Additionally, compared to SRCA using electrophoresis to analyze the amplification products, the visual closed-tube HNB-SRCA combined with the rapid DNA extraction protocol is a better potential cashew nut on-site detection strategy in foods.
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