Paralytic Shellfish Poisoning Detection by Surface Plasmon Resonance-Based Biosensors in Shellfish Matrixes

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• 作者：Eva S. Fonfrí ; a ; Natalia Vilariñ ; o ; Katrina Campbell ; Chris Elliott ; Simon A. Haughey ; Begoñ ; a Ben-Gigirey ; Juan M. Vieites ; Kentaro Kawatsu ; Luis M. Botana
• 刊名：Analytical Chemistry
• 出版年：2007
• 出版时间：August 15, 2007
• 年：2007
• 卷：79
• 期：16
• 页码：6303 - 6311
• 全文大小：274K
• 年卷期：v.79,no.16(August 15, 2007)
• ISSN：1520-6882

文摘

The detection of paralytic shellfish poisoning (PSP) toxinsin contaminated shellfish is essential for human healthpreservation. Ethical and technical reasons have promptedthe search for new detection procedures as an alternativeto the mouse bioassay. On the basis of the detection ofmolecular interactions by surface plasmon resonance(SPR) biosensors, an inhibition assay was developedusing an anti-GTX2/3 antibody (GT13-A) and a saxitoxin-CM5 chip. This assay allowed for quantification of saxitoxin (STX), decarbamoyl saxitoxin (dcSTX), gonyautoxin2,3 (GTX2/3), decarbamoyl gonyautoxin 2,3 (dcGTX2/3), gonyautoxin 5 (GTX5), and C 1,2 (C1/2) at concentrations from 2 to 50 ng/mL. The interference of five shellfishmatrixes with the inhibition assay was analyzed. Mussels,clams, cockles, scallops, and oysters were extracted withfive published methods. Ethanol extracts and acetic acid/heat extracts (AOAC Lawrence method) performed adequately in terms of surface regeneration and baselineinterference, did not inhibit antibody binding to the chipsurface significantly, and presented STX calibration curvessimilar to buffer controls in all matrixes tested. Hydrochloric acid/heat extracts (AOAC mouse bioassay method)presented surface regeneration problems, and althoughethanol-acetic acid/dichloromethane extracts performedwell, they were considered too laborious for routinesample testing. Overall the best results were obtained withthe ethanol extraction method with calibration curvesprepared in blank matrix extracts. STX recovery rate withthe ethanol extraction method was 60.52 ± 3.72%, withvariations among species. The performance of this biosensor assay in natural samples, compared to two AOACmethods for PSP toxin quantification (mouse bioassay andHPLC), suggests that this technology can be useful as aPSP screening assay. In summary, the GT13-A-STX chipinhibition assay is capable of PSP toxin detection inethanol shellfish extracts, with sufficient sensitivity toquantify the toxin in the range of the European regulatorylimit of 80 f">g/100 g of shellfish meat.