Full Name: Crustacean ELISA Kit (Tropomyosin)
Sample Type: Food (Bakery, Fish, Meat, Soups)
Sensitivity: 0.09 ppm
Crustacean form part of eight other food groups which can trigger allergies by the presence of immunoglobulin E antibodies. This usually leads to hypersensitivity reactions due to ingesting these crustaceans (examples include lobsters, shrimps and crabs), these are the most frequent causes of many food allergic reactions. Also, they are regarded as the third important foodstuff which can induce food related anaphylaxis. One of the major allergen found in shrimps and many other crustaceans is the ubiquitous muscle protein tropomyosin, this is also responsible for a large number of the ingestion related allergic reactions. This is because the heat of cooking destroys the allergenic tropomyosin from their bodies. In Japan, eating shrimps raw has no ill effect on those with allergies. The main allergen found in crabs is tropomyosin as well. Shellfish also are an important cause of many food allergic reactions by being able to trigger more than one type of allergy at once (examples include shellfish protein and histamine).
It is a highly conserved protein and is known to be homologous in a large number of different species (displaying considerable rate of identicalness). Tropomyosin is relatively resistant to heat and acidic digestion. The heat stability property allows this protein to be suitable for analysing in processed food samples. Some of the detection methods for food allergens (especially crustacean protein) need to be highly specific and be able to detect minute quantities of the allergen which could be present in complex food matrices. Currently, there are a few procedure available (PCR, lateral flow assays, ELISA) but not many are able to detect tropomyosin protein.
Crustacean ELISA kit is a procedure designed for detecting quantitative amounts of crustacean or tropomyosin in food products (such as fish, meat, bakery and soups). This assay has a minimum analytical sensitivity limit of 0.09 ppm.
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-tropomyosin antibodies.
- Tropomyosin Standards 1-5: Concentration 0, 20, 60, 200, 400 ppb.
- Conjugate (Anti-Tropomyosin-Peroxidase).
- Substrate Solution (TMB).
- Stop Solution.
- Extraction and sample dilution buffer (Tris)(10x Concentrate).
- Washing Solution (PBS + Tween 20)(10x).
- Instruction Manual.
The minimum detection sensitivity level of crustacean (tropomyosin) using current crustacean ELISA kit was 0.09 ppm. The standard range for this assay is 20.0 – 400.0 ppm.
– Sensitivity: Limit of detection, LOD (0.9ppm), Limit of quantification, LOQ (20.0ppm)
– Intra-Assay Precision: 6 – 8%
– Inter-Assay Precision: 5 – 12%
– Recovery: Meat (97%), Fish (93%), Vegetable soup (93%), Bakery products (90%), Soy sauce (84%).
– Linearity: 74 – 114%
– Cross Reactivity: The following displayed some cross reactions: Cockroach (protein) (0.01%). No cross reaction was detected for the following: Milk, Fish, Macadamia nut, Egg, Oyster, Chestnut, Wheat, Sunflower seeds, Pine nut, Rye, Pumpkin seeds, Soy, Oats, Cashew, Lecithin (soy), Barley, Peanut, Pea, Rice, Hazelnut, Bean, Corn, Almond, Potato, Buckwheat, Pecan, Carrot, Sesame, Coconut, Leek, Pork meat, Brazil nut, Celery, Beef, Walnut, Mustard, Chicken meat, Pistachio.
- A reliable and sensitive immunoassay for the determination of crustacean-protein in processed foods. J Agric Food Chem. (2007) 55 (23): 9345-50. Seiki K., et al.
- Molecular and immunological approaches in quantifying the air-borne food allergen tropomyosin in crab processing facilities. Int J Hyg Environ Health. (2014) 217 (7): 740-50. Kamath S.D., et al.
- Quantification of the major brown shrimp allergen Pen a 1 (tropomyosin) by a monoclonal antibody-based sandwich ELISA. J AllergyClin Immunol. (1997) 100 (2): 229-34. Jeoung B.J., et al.
- Identification of tropomyosinas the major shrimp allergen and characterization of its IgE-binding epitopes. J Immunol. (1993) 151 (10): 5354-63. Shanti K.N., et al.
- Seafood hypersensitivity in mite sensitized individuals: is tropomyosin the only responsible allergen? Ann Allergy Asthma Immunol. (2011) 106 (3): 223-9. Boquete M., et al.
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