title: An electrochemiluminescence-based competitive displacement immunoassay for the type-2 brevetoxins in oyster extracts.
abstract: A new competitive electrochemiluminescence-based immunoassay for the type-2 brevetoxins in oyster extracts was developed. The assay was verified by spiking known amounts of PbTx-3 into 80% methanol extracts of Gulf Coast oysters. We also provide preliminary data demonstrating that 100% acetone extracts, aqueous homogenates, and the clinical matrixes urine and serum can also be analyzed without significant matrix interferences. The assay offers the advantages of speed ( 2 h analysis time); simplicity (only 2 additions, one incubation period, and no wash steps before analysis); low limit of quantitation (conservatively, 50 pg/mL = 1 ng/g tissue equivalents); and a stable, nonradioactive label. Due to the variety of brevetoxin metabolites present and the lack of certified reference standards for liquid chromatography-mass spectrometry confirmation, a true validation of brevetoxins in shellfish extracts is not possible at this time. However, our assay correlated well with another brevetoxin immunoassay currently in use in the United States. We believe this assay could be useful as a regulatory screening tool and could support pharmacokinetic studies in animals and clinical evaluation of neurotoxic shellfish poisoning victims.
authors: United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, MD 21702-5011, USA. mark.poli@det.amedd.army.mil
authors: Mark A
authors: MA
authors: Poli
authors: Victor R
authors: VR
authors: Rivera
authors: Dwayne D
authors: DD
authors: Neal
authors: Daniel G
authors: DG
authors: Baden
authors: Shawn A
authors: SA
authors: Messer
authors: Steven M
authors: SM
authors: Plakas
authors: Robert W
authors: RW
authors: Dickey
authors: Kathleen El
authors: KE
authors: Said
authors: Leanne
authors: L
authors: Flewelling
authors: David
authors: D
authors: Green
authors: Jill
authors: J
authors: White
mesh_terms: D000818:Animals; D004563:Electrochemistry / Q000379:methods; D006801:Humans; D007118:Immunoassay / Q000379:methods; D049449:Luminescence; D008387:Marine Toxins / Q000097:blood / Q000737:chemistry* / Q000302:isolation & purification / Q000652:urine; D008958:Models, Molecular; D015394:Molecular Structure; D009498:Neurotoxins / Q000737:chemistry* / Q000302:isolation & purification; D010125:Ostreidae / Q000737:chemistry*; D035964:Oxocins / Q000097:blood / Q000737:chemistry* / Q000302:isolation & purification / Q000652:urine; D015203:Reproducibility of Results; D012428:Ruthenium; D014020:Tissue Extracts / Q000032:analysis*; D000095702:Polyether Toxins
chemical_list: D008387:Marine Toxins; D009498:Neurotoxins; D035964:Oxocins; D012428:Ruthenium; D014020:Tissue Extracts; C053342:brevetoxin; D000095702:Polyether Toxins