Fish ELISA Kit

Full Name: Fish ELISA Kit (Parvalbumin)
Sample Type: Food (Asian Food, Crackers, Sauces, Soups, Surimi, Wine)
Sensitivity: 1.4 ppm

BACKGROUND

Fish is a vital source of dietary protein and in many places it represent the main component of the diet, especially near coastal areas. However, allergy to fish is a prevalent cause for a large number of food allergic reactions. One of the major allergen is parvalbumin, this a 12 kDa protein and it is about 108 amino acids in length. Fish belongs to the class of animals known as cartilaginous fish and its protein is composed of caseins, globulins, myosins, actin and tropomyosin. The percentage of fat in the fish protein is about 1%.

The following assay offers a highly sensitive detection method for fish that is based on the parvalbumin allergen. It can be used to quantify fish-residues present in soups, wine, crackers, surimi, sauces and many products used in the Asian culture. The abundant parvalbumin was one of the first food allergens to be isolated, crystallised and then identified on a molecular level. They are sarcoplasmatic muscle proteins which are highly water-soluble, well conserved and are resistant to extreme pH, denaturing agents and heat.

INTENDED USE

Fish ELISA kit is designed for detecting quantitative levels of fish (in particular allergen parvalbumin) in many different foods (such as sauces, soups, crackers, wine, Asian food and surimi). This assay has a minimum analytical sensitivity limit of 1.4 ppm.

CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

• Microtiter Plate: Coated with anti-fish antibodies.
• Cod Standards 1-5: Concentration 0, 4, 10. 40, 100 ppm.
• Conjugate (Anti-Fish-Peroxidase).
• Substrate Solution (TMB).
• Stop Solution.
• Extraction and sample dilution buffer (Tris)(10x Concentrate).
• Washing Solution (10x).
• Instruction Manual.

SENSITIVITY

The minimum detection sensitivity level of fish or parvalbumin allergen’s using current fish ELISA kit was 1.4 ppm. The standard range for this assay is 4.0 – 100.0 ppm.

ASSAY CHARACTERISTICS

– Sensitivity: Limit of detection, LOD (1.4ppm), Limit of quantification, LOQ (4.0ppm)
– Intra-Assay Precision: 7 – 12%
– Inter-Assay Precision: 4 – 10%
– Linearity: 89 – 105%
– Specificity: No cross reaction was detected for the following: Almond, Hazelnut, Pork, Barley, Isinglass, Potato, Bean, Lamb, Pumpkin seed, Beef, Macadamia, Rice, Brazil nut, Milk, Rye, Buckwheat, Millet, Scallop, Carrot, Mustard, Sesame, Cashew, Oat, Shrimp, Celery, Onion, Soy, Chicken, Pea, Sunflower seed, Corn, Peanut, Walnut, Egg, Pecan, Wheat, Fish gelatin, Pistachio.
– Recovery: Soup (117%), Surimi (114%), Worcester Sauce (112%), Wine (red)(103%), Asia Sauce (103%), Cracker (99%), Spring Roll (93%).

REFERENCES

1. Immunological cross-reactivity between four distant parvalbumins-Impact on allergen detection and diagnostics. Mol Immunol. (2015) 63 (2): 437-48. Sharp M.F., et al.
2. A sensitive enzyme-linked immunosorbent assay for the determination of fish-protein in processed foods. Food Chem. (2013) 136 (2): 675-81. Shibahara Y., et al.
3. Monoclonal antibody specific to a major fish allergen: parvalbumin. J Food Prot. (2009) 72 (4): 818-25. Gajewski K.G. and Hsieh Y.H.
4. Quantitative sandwich ELISA for the determination of fish in foods. J Immunol Methods. (2008) 329 (1-2): 45-55. Faeste C.K. and Plassen C.
5. Protein carbonylation during electron beam irradiation may be responsible for changes in IgE binding to turbot parvalbumin. Food Chem Toxicol. (2014) 69: 32-7. Li Z., et al.
6. Parvalbumin–the major tropical fish allergen. Pediatr Allergy Immunol. (2008) 19 (5): 399-407. Lim D.L., et al.

OTHER RELATED ELISA KITS

• Crustaceans (Tropomyosin) ELISA Kit
• Ovalbumin ELISA Kit
• Mollusc (Tropomyosin) ELISA Kit