title: Detection of paralytic shellfish toxins by a solid-phase inhibition immunoassay using a microsphere-flow cytometry system.
abstract: Paralytic shellfish poisoning is a toxic syndrome described in humans following the ingestion of seafood contaminated with saxitoxin and/or its derivatives. The presence of these toxins in shellfish is considered an important health threat and their levels in seafood destined to human consumption are regulated in many countries, as well as the levels of other chemically unrelated toxins. We studied the feasibility of immunodetection of saxitoxin and its analogs using a solid-phase microsphere assay coupled to flow cytometry detection in a Luminex 200 system. The technique consists of a competition assay where the toxins in solution compete with bead-bound saxitoxin for binding to an antigonyautoxin 2/3 monoclonal antibody (GT-13A). The assay allowed the detection of saxitoxin both in buffer and mussel extracts in the range of 2.2-19.7 ng/mL (IC(20)-IC(80)). Moreover, the assay cross-reactivity with other toxins of the group is similar to previously published immunoassays, with adequate detection of most analogs except N-1 hydroxy analogs. The recovery rate of the assay for saxitoxin was close to 100%. This microsphere-based immunoassay is suitable to be used as a screening method, detecting saxitoxin from 260 to 2360 μg/kg. This microsphere/flow cytometry system provided similar sensitivities to previously published immunoassays and provides a solid background for the development of easy, flexible multiplexing of toxin detection in one sample.
authors.affiliation: Departamento de Farmacología, Facultad de Veterinaria, Universidad de Santiago de Compostela, 27002 Lugo, Spain.
authors.forename: María
authors.initials: M
authors.lastname: Fraga
authors.forename: Natalia
authors.initials: N
authors.lastname: Vilariño
authors.forename: M Carmen
authors.initials: MC
authors.lastname: Louzao
authors.forename: Katrina
authors.initials: K
authors.lastname: Campbell
authors.forename: Christopher T
authors.initials: CT
authors.lastname: Elliott
authors.forename: Kentaro
authors.initials: K
authors.lastname: Kawatsu
authors.forename: Mercedes R
authors.initials: MR
authors.lastname: Vieytes
authors.forename: Luis M
authors.initials: LM
authors.lastname: Botana
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D049872:Bivalvia; D005434:Flow Cytometry; D059022:Food Safety / Q000379:methods*; D006801:Humans; D007118:Immunoassay; D008863:Microspheres; D012530:Saxitoxin / Q000031:analogs & derivatives / Q000032:analysis* / Q000276:immunology; D057096:Shellfish Poisoning
chemical_list: D000911:Antibodies, Monoclonal; D012530:Saxitoxin