Okatest

HOW DOES IT HELP YOU?

Okatest can help you to choose the best area and time to harvest, preventing extraction of contaminated molluscs from areas near those ones closed due to presence of these toxins.
Performing shelf-control with Okatest before harvesting or as end-product testing you can assure molluscs are fit for consumption and avoid unnecessary process of contaminated products.
Okatest allows you to deliver results on the same day as you can process up to 43 samples in about 3 hours. In addition, you will be able to combine it with the reference method to get results for the whole group of lipophilic toxins in less time; as well as to follow up areas closed by okadaic acid.

OkaTest can therefore be used as a supplementary test to the reference method for determination of the OA-toxins group in molluscs according to the Commission Regulations (EC) No. 853/2004 and No. 15/2011 (http://www.aecosan.msssi.gob.es/en/CRLMB/web/public_documents/seccion/other_crlmb_standard_operating_procedures.htm)

ASSAY PROCEDURE

TEST PRINCIPLE

Inhibition of serine/threonine protein phosphatases (PP1 & PP2A) activity by this group of toxins is assumed to be responsible for these diarrheic disorders. Okatest based on the phophatase activity inhibition (PP2A) by OA-toxins group, responsible for diarrheic shellfish poisoning (DSP).
Phosphatase enzyme PP2A is able to hydrolyse a specific substrate, yielding a product that can be detected at 405 nm. Samples containing toxins from the okadaic acid group will inhibit the enzyme activity proportionally to the amount of toxin contained in the sample.

TECHNICAL DATA

• Working range: 63 -352 μg OA equivalents/Kg.
• Assay time: 1 h extraction plus 2 steps of 20 & 30 minutes.
• Presentation: 96 and 48-well microtiter plate. Total of 43 samples.
• Equipment: Microtiter plate reader at 405 nm.

Validations:

Intralaboratory validation:
Smienk et al. 2012 Single Laboratory Validation of A Ready-to-Use Phosphatase Inhibition Assay for Detection of Okadaic Acid Toxins. Toxins, 5, 339-352.

Inter-laboratory validadation:

Validación con 16 laboratorios procedentes de 11 países. Smienk et al. 2013. Quantitative Determination of the Okadaic Acid Toxins Group by a Colorimetric Phosphatase Inhibition Assay: Interlaboratory Study. Journal of AOAC International Vol. 96, No. 1.

Participation in interlaboratory studies: Quasimeme laboratory performance studies 2012-2015

Legislation:

Regulation (EC) No 853/2004 of the European Parliament and of the Council of 29 April 2004 laying down specific hygiene rules for food of animal origin.

Commission Regulation (EC) No 2074/2005 of 5 December 2005 laying down implementing measures for certain products under Regulation (EC) No 853/2004 of the European Parliament and of the Council and for the organisation of official controls under Regulation (EC) No 854/2004 of the European Parliament and of the Council and Regulation (EC) No 882/2004 of the European Parliament and of the Council, derogating from Regulation (EC) No 852/2004 of the European Parliament and of the Council and amending Regulations (EC) No 853/2004 and (EC) No 854/2004.

Commission Regulation (EU) No 15/2011 of 10 January 2011 amending Regulation (EC) No 2074/2005 as regards recognised testing methods for detecting marine biotoxins in live bivalve molluscs.

Related articles