Fish Parvalbumin ELISA Kit

Sample

wine, soups, sauces, crackers, surimi and asia products

Species Reactivity

Fish

Detection Method

sELISA

Intended Use

The Fish Parvalbumin ELISA Kit represents a highly sensitive detection system for fish, based on the trans-species allergen parvalbumin. It is particularly capable of the quantification of fish residues in wine, soups, sauces, crackers, surimi and asia products.

Contents of Kit

1. Microtiter plate consisting of 12 strips with 8 breakable wells each, coated with anti-fish antibodies.
2. Cod Standards (0; 4; 10; 40; 100 ppm of cod): 5 vials with 2.0 mL each, dyed red, ready-to-use.
3. Conjugate (anti-fish-peroxidase): 15 mL, dyed red, ready-to-use.
4. Substrate Solution (TMB): 15 mL, ready-to-use.
5. Stop Solution (0.5 M H₂SO₄): 15 mL, ready-to-use.
6. Extraction and sample dilution buffer (Tris): 2×120 mL as 10× concentrate, dyed red. Dilute 1+9 with distilled water. Stored at 4°C the diluted buffer is stable for at least one week. If during the cold storage crystals precipitate, the concentrate should be warmed up to 37°C for 15 minutes.
7. Washing Solution (PBS + Tween 20): 60 mL as 10× concentrate. Dilute 1+9 with distilled water. Stored at 4°C the diluted buffer is stable for at least 4 weeks. If during the cold storage crystals precipitate, the concentrate should be warmed up to 37°C for 15 minutes.
8. Plastic bag to store unused microtiter strips.
9. Instruction Manual.

Storage

The kit contains reagents for 96 determinations. They have to be stored at 2-8°C. Expiry data are printed on the labels of the bottles and the outer package.

Precision

Intra-assay Precision 7-12%
Inter-assay Precision 4-10%

Sensitivity

The limit of detection (LOD) of the Fish Parvalbumin ELISA Kit is 1.4 ppm (cod) for the standard curve.
Validation experiments with common matrices resulted in the following LODs [ppm].

The limit of quantification (LOQ) of the Fish Parvalbumin ELISA Kit is 4 ppm.
Due to the variety of sample matrices and their influence on the blank, results less than the LOQ should be treated as negative.

General Description

Fishes belong to the most frequent elicitors of food allergies. The allergies are predominantly induced by the low-molecular, calcium-binding muscle protein parvalbumin. The protein is characterized by its high heat resistance and stability against denaturing agents and proteolytic enzymes. Predominantly in regions with a high consumption of fish like Scandinavia, Japan or the Mediterranean countries, fish allergies represent a heavy health problem. The symptoms are ranging from inflammation of the skin over gastrointestinal and respiratory problems up to live-threatening anaphylactic shock. In spite of the high biodiversity most patients react with allergic symptoms to several fish species due to the high cross-reactivity between the fish allergens. For fish-allergic persons hidden fish allergens in food are a critical problem. Already very low amounts of fish can cause allergic reactions, which may lead to anaphylactic shock in severe cases. Because of this, fish-allergic persons must strictly avoid the consumption of fish containing food. Cross-contamination, mostly in consequence of the production process, is often noticed. This explains why in many cases the existence of fish residues in food cannot be excluded. For this reason sensitive detection systems for fish residues in foodstuffs are required.

Standard Curve

The following table contains an example for a typical standard curve. The binding is calculated as percent of the absorption of the 100 ppm standard. These values are only an example and should not be used instead of the standard curve which has to be measured in each new test.