Monoclonal Antibody‐Based Sandwich ELISA for Reliable Identification of Imported Pangasius Catfish

Abstract

Pangasius catfish, primarily tra (Pangasius hypophthalmus) and basa (Pangasius bocourti), are farm-raised catfish imported from Asia and have become a common substitute for domestic catfish, grouper, and other high-valued fish in restaurant-served dishes in the United States. This article reports on the development of a monoclonal antibody-based sandwich enzyme-linked immunosorbent assay (ELISA) for the identification of cooked Pangasius fish, basa, and tra. The assay uses a pair of monoclonal antibodies (MAbs F7B8 and T7E10) specific to a heat-stable 36-kDa protein present in a saline extraction of the fish muscle; MAb F7B8, which cross-reacts to all fish species, is the capture antibody and the biotin-conjugated MAb T7E10, which is specific to Pangasius fish, is the detection antibody. This sandwich ELISA reliably identified fully cooked basa and tra from more than 70 common finfish, shellfish, land animal species, and other protein materials tested. It can also sensitively detect 0.5% of adulterated basa or 0.1% tra in a mixed crabmeat products with low intra-assay (%CV: 2.59 to 4.14) and inter-assay (%CV: 3.36 to 3.71) variabilities. The new assay provides a rapid and reliable means of distinguishing fish in the Pangasiidae family from other common food fish and nonfish species and will greatly assist efforts to discourage the illegal practice of substituting high-value popular fish species by the cheaper farm-raised imported Pangasius fish at the retail and restaurant levels.

Bushmeat Species Identification: Recombinase Polymerase Amplification (RPA) Combined with Lateral Flow (LF) Strip for Identification of Formosan Reeves’ Muntjac (Muntiacus reevesi micrurus)

“…A variety of techniques analyzing protein composition have been utilized to identify meat origin [1,5,6,30]. However, heat and pressure processing may cause protein denaturation, posing problems of possible misdiagnosis [31].…”

“…A variety of analytic methods have been applied to differentiate animal species of meat, including ELISA [1][2][3][4], immunochromatography [5], and Raman spectroscopy [6,7]. Currently, PCR-based techniques, such as species-specific PCR [8][9][10][11][12], restriction fragment length polymorphism (RFLP) [13,14], and real-time PCR [15][16][17], are widely used.…”

Rapid Immune Colloidal Gold Strip for Cetacean Meat Restraining Illegal Trade and Consumption: Implications for Conservation and Public Health

“…Moreover, it is likely that the 2 mAbs in this study would not inhibit each other’s binding to the antigen because these 2 mAbs were developed using epitopes with known relative locations. Therefore the following sandwich-based immunoassay that is the most commonly used format in commercial kits [24] could be applied. In contrast, it is needed to do epitope comparison by performing additivity test for identifying a suitable pair of mAbs before designing a sandwich-based immunoassay when the mAbs are raised from purified protein or crude protein extract and the information of epitopes is unknown.…”

Microarray Chip Development Using Infrared Imaging for the Identification of Catfish Species

“…Due to their commercial importance, a few research groups have focused on developing methods for the specific detection of select catfish species. A monoclonal antibody-based sandwich enzyme-linked immunosorbent assay (ELISA) was developed to identify different species of Pangasius , 14 and both Wong et al 15 and Bhattacharjee et al 16 looked more in depth at the use of DNA barcoding to identify catfish specifically.…”