Multidetection of Paralytic, Diarrheic, and Amnesic Shellfish Toxins by an Inhibition Immunoassay Using a Microsphere-Flow Cytometry System
- 31 July 2013
- journal article
- research article
- Published by American Chemical Society (ACS) in Analytical Chemistry
- Vol. 85 (16), 7794-7802
- https://doi.org/10.1021/ac401146m
Abstract
The presence of paralytic shellfish poisoning (PSP), diarrheic shellfish poisoning (DSP), and amnesic shellfish poisoning (ASP) toxins in seafood is a severe and growing threat to human health. In order to minimize the risks of human exposure, the maximum content of these toxins in seafood has been limited by legal regulations worldwide. The regulated limits are established in equivalents of the main representatives of the groups: saxitoxin (STX), okadaic acid (OA), and domoic acid (DA), for PSP, DSP, and ASP, respectively. In this study a multidetection method to screen shellfish samples for the presence of these toxins simultaneously was developed. Multiplexing was achieved using a solid-phase microsphere assay coupled to flow-fluorimetry detection, based on the Luminex xMap technology. The multidetection method consists of three simultaneous competition immunoassays. Free toxins in solution compete with STX, OA, or DA immobilized on the surface of three different classes of microspheres for binding to specific monoclonal antibodies. The IC50 obtained in the buffer was similar in single- and multidetection: 5.6 +/- 1.1 ng/mL for STX, 1.1 +/- 0.03 ng/mL for OA, and 1.9 +/- 0.1 ng/mL for DA. The sample preparation protocol was optimized for the simultaneous extraction of STX, OA, and DA with a mixture of methanol and acetate buffer. The three immunoassays performed well with mussel and scallop matrixes displaying adequate dynamic ranges and recovery rates (around 90% for STX, 80% for OA, and 100% for DA). This microsphere-based multidetection immunoassay provides an easy and rapid screening method capable of detecting simultaneously in the same sample three regulated groups of marine toxins.Keywords
This publication has 38 references indexed in Scilit:
- Development of a rapid low cost fluorescent biosensor for the detection of food contaminantsBiosensors and Bioelectronics, 2013
- Development of a Solid-Phase Receptor-Based Assay for the Detection of Cyclic Imines Using a Microsphere-Flow Cytometry SystemAnalytical Chemistry, 2013
- Effect of Uncontrolled Factors in a Validated Liquid Chromatography–Tandem Mass Spectrometry Method Question Its Use As a Reference Method for Marine Toxins: Major Causes for ConcernAnalytical Chemistry, 2011
- Development of a multiplex flow cytometric microsphere immunoassay for mycotoxins and evaluation of its application in feedMycotoxin Research, 2010
- Neurotoxic Alkaloids: Saxitoxin and Its AnalogsMarine Drugs, 2010
- Development of ELISAs for detecting domoic acid, okadaic acid, and saxitoxin and their applicability for the detection of marine toxins in samples collected in BelgiumFood Additives & Contaminants: Part A, 2010
- Single Laboratory Validation of a Surface Plasmon Resonance Biosensor Screening method for Paralytic Shellfish Poisoning ToxinsAnalytical Chemistry, 2010
- Antibody-Based Protein Multiplex Platforms: Technical and Operational ChallengesClinical Chemistry, 2010
- Assessment of Specific Binding Proteins Suitable for the Detection of Paralytic Shellfish Poisons Using Optical Biosensor TechnologyAnalytical Chemistry, 2007
- Domoic Acid Levels of Naturally Contaminated Scallops as Affected by CanningJournal of Food Science, 1998