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[A indirect competitive ELISA to detect domoic acid in seawater and shellfish]
Renyan Liu1, Daoyan Xu, Yuhua Dong
1National Marine Environmental Monitoring Center, Dalian 116023, China. ryliu@nmemc.gov.cn
Objective:
A competitive enzyme-linked immunosorbent assay (ELISA) was developed for measurent domoic acid (DA) in seawater and shellfish.
Methods:
DA was coupled to bovine serum albumin (BSA) and ovalbumin (OVA) by carbodiimide reaction. DA-BSA as immunogen was injected in BALB/c mice. Titres of the antisera against DA were determinated using DA-OVA as coating ligand by ELISA method. After female BALB/c mice were immunized six times, the polyclonal antibodies anti-DA was obtained. The competitive indirect ELISA for domoic acid in shellfish and seawater was established.
Results:
Under optimal condition, the detection limit of DA was 10.0 ng/ml (equal 4 microg/g shellfish meat). The recovery of domoic acid added in seawater was 83.2% - 124.7% with a coefficient of variation of 4.7% - 5.9%, the recovery of domoic acid added in shellfish was 85.9% - 99.9% with a coefficient of variation of 2.4% - 7.1%.
