Development of ELISAs for detecting domoic acid, okadaic acid, and saxitoxin and their applicability for the detection of marine toxins in samples collected in Belgium.
Okadaic acid, a diarrhetic shellfish poison, domoic acid, an amnesic shellfish poison, and saxitoxin, a paralytic shellfish poison, are three of the best-known marine biotoxins. The mouse bioassay is the method most widely used to detect many of these toxins in shellfish samples, but animal welfare concerns have prompted researchers to seek alternative methods of detection. In this study, three direct competitive enzyme-linked immunosorbent assays (ELISAs), each based on antibodies raised in rabbits against a conjugate of the analyte of interest, were developed for marine biotoxin detection in mussel, oyster, and scallop. One assay was for okadaic acid, one for saxitoxin, and one for domoic acid usually detected and quantified by high-performance liquid chromatography-ultraviolet light (HPLC-UV). All three compounds and a number of related toxins were extracted quickly and simply from the shellfish matrices with a 9 : 1 mixture of ethanol and water before analysis. The detection capabilities (CCbeta values) of the developed ELISAs were 150 microg kg(-1) for okadaic acid, 50 microg kg(-1) for domoic acid, and 5 microg kg(-1) or less for saxitoxin. The assays proved satisfactory when used over a 4-month period for the analysis of 110 real samples collected in Belgium.
Dubois
M
M
Departement Sante, CERgroupe, B-6900 Marloie, Belgium. m.dubois@cergroupe.be
Demoulin
L
L
Charlier
C
C
Singh
G
G
Godefroy
S B
SB
Campbell
K
K
Elliott
C T
CT
Delahaut
Ph
P
D000818:Animals; D000918:Antibody Specificity; D001530:Belgium; D049872:Bivalvia / Q000737:chemistry*; D002138:Calibration; D002273:Carcinogens / Q000032:analysis; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D007608:Kainic Acid / Q000031:analogs & derivatives* / Q000032:analysis; D008387:Marine Toxins / Q000032:analysis; D051379:Mice; D019319:Okadaic Acid / Q000032:analysis*; D010125:Ostreidae / Q000737:chemistry*; D049897:Pectinidae / Q000737:chemistry*; D011817:Rabbits; D012530:Saxitoxin / Q000032:analysis*; D012680:Sensitivity and Specificity
D002273:Carcinogens; D008387:Marine Toxins; D019319:Okadaic Acid; D012530:Saxitoxin; C012301:domoic acid; D007608:Kainic Acid