Simultaneous detection of five potentially allergenic tropical fruit species (mango, pineapple, avocado, banana, and coconut) using multiplex polymerase chain reaction coupled with capillary electrophoresis

In this study, a multiplex polymerase chain reaction (PCR) assay that can simultaneously and sensitively detect the presence of five potentially allergenic tropical fruit species was developed to prevent allergic reactions among consumers and enhance future regulations on food allergy labeling. The novel species-specific primer set was designed with high specificity and sensitivity so that it can be applied even in processed foods. The primer set targets chloroplast genes matK and ITS regions. The obtained amplicon sizes were 114 bp for mango, 132 bp for pineapple, 158 bp for avocado, 183 bp for banana, and 242 bp for coconut. The specificity of the multiplex PCR assay was validated with 27 different plant species including five tropical fruit species, demonstrating no cross-reactivity and confirming detection sensitivity at approximately 1 pg. Its applicability was further verified using 32 commercially available food products. In particular, juice products containing two distinct fruit types displayed amplification signals for both species, highlighting the method's ability to detect multiple species simultaneously.