Identification of Grouper (Epinephelus guaza), Wreck Fish (Polyprion americanus), and Nile Perch (Lates niloticus) Fillets by Polyclonal Antibody-Based Enzyme-Linked Immunosorbent Assay

Abstract

An indirect enzyme-linked immunosorbent assay (ELISA) has been developed for the species identification of grouper (Epinephelus guaza), wreck fish (Polyprion americanus), and Nile perch (Lates niloticus) fillets. The assay was performed in two different formats, microtiter plates and immunostick tubes, and uses polyclonal antibodies raised in rabbits against muscle-soluble proteins of grouper (anti-GSP), wreck fish (anti-WSP), and Nile perch (anti-PSP). The antibodies were made species-specific by blocking them with the heterologous soluble muscle proteins. Immunorecognition of polyclonal antibodies adsorbed to their specific fish samples was made with swine antirabbit immunoglobulins conjugated to the enzyme horseradish peroxidase. Subsequent enzymatic conversion of the substrate allowed unequivocal identification of the species studied.

Improved fish species identification by use of lab-on-a-chip technology

“…In terms of simplicity and speed, antibody based methods would be most appropriate. However, only a limited number of immunoassays have been developed and none are available for wide scale commercial use (Asensio et al, 2003;Cespedes et al, 1999a). DNA based methods have, therefore, become the most appropriate approach to species identification as DNA remains detectable in all but the most heavily processed samples.…”

Examination of Meat Components in Commercial Dog and Cat Food by Using Polymerase Chain Reaction-Restriction Fragment Length Polymorphisms (PCR-RFLPs) Technique

“…There are several techniques of analysis of food component, including mid-infrared spectroscopy, enzyme-linked immunosorbent assay, and capillary electrophoresis. These techniques are expensive, time consuming, and/or of low specificity [1,2,8,9]. Usually, DNA samples are more stable than proteins during thermal treatment.…”

High-Throughput Proteomics: A New Tool for Quality and Safety in Fishery Products

“…Studies using gelbased proteomics and immunological methodologies have been extensively applied for the authentication of fish species [42][43][44] but one of the first gel-free proteomic strategies developed in fish was used by Mazzeo and co-workers [45] who developed a top-down method based on matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) MS. The authors were able to discriminate 25 different commercial fish species by selecting sarcoplasmic proteins with molecular weight about 11KDa as species-specific biomarkers for their unambiguous identification.…”