Single-Laboratory Validation of the Biosense Direct Competitive Enzyme-Linked Immunosorbent Assay (ELISA) for Determination of Domoic Acid Toxins in Shellfish

Method validation was conducted for an enzyme-linked immunosorbent assay (ELISA) for the determination of domoic acid (DA) toxins, known to give amnesic shellfish poisoning (ASP) symptoms, in shellfish. The calibration curve range of the assay is approximately 10-260 pg/mL, with a dynamic working ra...

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Bibliographic Details
Published in:Journal of AOAC International 2007-07, Vol.90 (4), p.1000-1010
Main Authors: Kleivdal, H, Kristiansen, S.I, Nilsen, M.V, Briggs, L
Format: Article
Language:English
Subjects:
amnesic shellfish poisoning
Animals
Biological and medical sciences
Biosensing Techniques
biosensors
Bivalvia - metabolism
Calibration
Chemistry Techniques, Analytical - methods
Chemistry, Pharmaceutical - methods
detection
domoic acid
Dose-Response Relationship, Drug
Enzyme-linked immunosorbent assay
Enzyme-Linked Immunosorbent Assay - methods
Fish and seafood industries
Food Analysis
Food Contamination
Food industries
Fundamental and applied biological sciences. Psychology
Identification and classification
Kainic Acid - analogs & derivatives
Kainic Acid - chemistry
laboratory techniques
Marine toxins
Marine Toxins - chemistry
Methods
mussels
Neurotoxic agents
oysters
Pectinidae - metabolism
Reproducibility of Results
scallops
seafoods
Shellfish
toxic substances
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Summary:Method validation was conducted for an enzyme-linked immunosorbent assay (ELISA) for the determination of domoic acid (DA) toxins, known to give amnesic shellfish poisoning (ASP) symptoms, in shellfish. The calibration curve range of the assay is approximately 10-260 pg/mL, with a dynamic working range for DA toxins in shellfish from 0.01 to at least 250 mg/kg. The ASP ELISA showed no significant cross-reactivity to structural analogs, and proved to be robust to deliberate alterations of the optimal running conditions. The shellfish matrix effects observed with mussels, oysters, and scallops were eliminated by diluting shellfish extracts 1:200 prior to analysis, leading to a limit of detection at 0.003 mg/kg. Thirteen blank shellfish homogenates were spiked with certified mussel material containing DA to levels in the range of 0.1-25 mg DA/kg, and analyzed in quadruplicate on 3 different days. The relative standard deviation (RSD) under intra-assay repeatability conditions ranged from 6.5 to 13.1%, and under interassay repeatability conditions the RSD ranged from 5.7 to 13.4%, with a mean value of 9.3%. The recoveries ranged from 85.5 to 106.6%, with a mean recovery of 102.2%. A method comparison was conducted with liquid chromatography with ultraviolet detection, using naturally contaminated scallop samples (n = 27) with DA levels at 0-244 mg/kg. The overall correlation coefficient was 0.960 and the slope of the regression was 1.218, indicating a good agreement between the methods.
ISSN:1060-3271
1944-7922
DOI:10.1093/jaoac/90.4.1000