RESUMEN
Food allergies have become a global issue and are estimated to affect approximately 220 million people worldwide. Allergy to peanuts can easily become life-threatening and induce anaphylactic reactions. Mislabeling and cross-contamination during food processing can occur in the frame of world population growth and pose a serious health issue. As the mandatory allergen list is not uniform worldwide, the development of routine analytical strategies with high specificity and sensitivity is a demanding task to aid in the rapid identification of allergenic foods. In this work, an electrochemical aptasensor for Ara h1 peanut allergen was developed by immobilizing the specific aptamer by the inserting method. First, a layer of p-aminothiophenol (p-ATP) was immobilized on the gold surface of screen-printed electrodes (GSPE) to improve the aptamer insertion and reduce the fouling effects at the electrode surface. The grafting of the p-ATP and Ara h1 aptamer on the GSPE surface was monitored by cyclic voltammetry (CV), differential pulse voltammetry (DPV), and electrochemical impedance spectroscopy (EIS). The resulting disposable aptasensor allowed for indirect electrochemical detection of Ara h1 protein in the presence of 5 mM ferro/ferricyanide as a redox probe. The electrochemical response upon aptamer-target interaction was monitored in the concentration range 1-250 nM, and two limits of detection in the nanomolar range were estimated based on DPV (2.78 nM Ara h1) and EIS (0.82 nM Ara h1) measurements. The aptasensor was successfully applied to real sample analysis.
Asunto(s)
Compuestos de Anilina , Incrustaciones Biológicas , Hipersensibilidad a los Alimentos , Compuestos de Sulfhidrilo , Humanos , ADN , Oligonucleótidos , Arachis , Oro , Alérgenos , Adenosina TrifosfatoRESUMEN
This review summarizes the stepwise strategy and key points for magnetic beads (MBs)-based aptamer selection which is suitable for isolating aptamers against small and large molecules via systematic evolution of ligands by exponential enrichment (SELEX). Particularities, if any, are discussed according to the target size. Examples targeting small molecules (<1000 Da) such as xenobiotics, toxins, pesticides, herbicides, illegal additives, hormones, and large targets such as proteins (biomarkers, pathogens) are discussed and presented in tabular formats. Of special interest are the latest advances in more efficient alternatives, which are based on novel instrumentation, materials or microelectronics, such as fluorescence MBs-SELEX or microfluidic chip system-assisted MBs-SELEX. Limitations and perspectives of MBs-SELEX are also reviewed. Taken together, this review aims to provide practical insights into MBs-SELEX technologies and their ability to screen multiple potential aptamers against targets from small to large molecules.
Asunto(s)
Herbicidas , Cromatografía de Afinidad , Ligandos , Microfluídica , OligonucleótidosRESUMEN
Food contaminants represent possible threats to humans and animals as severe food safety hazards. Prolonged exposure to contaminated food often leads to chronic diseases such as cancer, kidney or liver failure, immunosuppression, or genotoxicity. Aflatoxins are naturally produced by strains of the fungi species Aspergillus, which is one of the most critical and poisonous food contaminants worldwide. Given the high percentage of contaminated food products, traditional detection methods often prove inadequate. Thus, it becomes imperative to develop fast, accurate, and easy-to-use analytical methods to enable safe food products and good practices policies. Focusing on the recent progress (2018-2023) of electrochemical aptasensors for aflatoxin B1 (AFB1) detection in food and beverage samples, without pretending to be exhaustive, we present an overview of the most important label-free and labeled sensing strategies. Simultaneous and competitive aptamer-based strategies are also discussed. The aptasensors are summarized in tabular format according to the detection mode. Sample treatments performed prior analysis are discussed. Emphasis was placed on the nanomaterials used in the aptasensors' design for aptamer-tailored immobilization and/or signal amplification. The advantages and limitations of AFB1 electrochemical aptasensors for field detection are presented.