An electrochemiluminescence-based competitive displacement immunoassay for the type-2 brevetoxins in oyster extracts.
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.
Poli
Mark A
MA
United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, MD 21702-5011, USA. mark.poli@det.amedd.army.mil
Rivera
Victor R
VR
Neal
Dwayne D
DD
Baden
Daniel G
DG
Messer
Shawn A
SA
Plakas
Steven M
SM
Dickey
Robert W
RW
Said
Kathleen El
KE
Flewelling
Leanne
L
Green
David
D
White
Jill
J
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
D008387:Marine Toxins; D009498:Neurotoxins; D035964:Oxocins; D012428:Ruthenium; D014020:Tissue Extracts; C053342:brevetoxin; D000095702:Polyether Toxins