Filling Analytical Gaps in Allergen Detection─Real-Time PCR for the Detection of Commercially Relevant Cephalopods and Gastropods in Food

Abstract

Mollusks belong to the group of shellfish, which are considered to be among the elicitors of severe food allergies worldwide. In recent years, numerous PCR detection methods have been developed for other shellfish such as crustaceans. However, cephalopods and gastropods were not considered in the development of these shellfish detection systems. In this study, we have developed highly specific real-time PCR methods for the comprehensive detection of all commercially relevant cephalopod species and the gastropod families Helicidae, Buccinidae, and Muricidae in food matrices. In total, we cross-tested over 100 animal and plant species to show the specificity of our systems. The limit of detection (LOD 12 ) was set at 1 pg of cephalopod and gastropod DNA or 10 ppm (mg/kg) spiked in a vegetarian food product. The robustness of the protocol was confirmed by testing multiple parameters while cooking and autoclaving of samples ensured the practical applicability of the systems.

Ratiometric Fluorescence Aptasensor of Allergen Protein Based on Multivalent Aptamer-Encoded DNA Flowers as Fluorescence Resonance Energy Transfer Platform

“…Up to now, a series of analysis strategies have been established for the qualitative or quantitative analysis of Ara h1, such as polymerase chain reaction (PCR), ,, loop-mediated isothermal amplification (LAMP), , liquid chromatography–tandem mass spectrometry (LC–MS/MS), fluorescence assay, surface-enhanced Raman spectroscopy (SERS), enzyme-linked immunosorbent assay (ELISA), electrochemical method, lateral flow assay (LFA), and surface plasmon resonance (SPR) . Fluorescence-based biosensor platforms, in particular, exhibit significant application potential owing to their simplicity, rapidity, and high sensitivity. , Nevertheless, the majority of fluorescence strategies are typically assessed at a single emission wavelength, posing challenges in mitigating autofluorescence interference in intricate matrices of food samples .…”