AMOZ ELISA Kit

Sample

Milk, Honey, Egg, Egg powder, Shrimps, Tissue/fish, Liver, Urine

Detection Method

cELISA

Intended Use

A competitive enzyme immunoassay for screening and quantitative analysis of AMOZ.

Contents of Kit

1. One sealed (96-wells) microtiter plate (12 strips, 8 wells each), coated antibody. Ready-to-use.
2. Position of the reagents in the kit. For preparation of the reagents see Reagent Preparation.
3. Sample dilution buffer (40 ml, ready-to-use)
4. Rinsing buffer (30 ml, 20× concentrated)
5. Substrate solution (12 ml, ready-to-use)
6. Stop solution (15 ml, ready-to-use)
7. Conjugate solution (100 μl, 100 times concentrated)
8. Dilution buffer (15 ml, ready-to-use)
9. AMOZ-stock solution (1 ml 100 ng/ml for spiking samples )
10. Zero Standard (2ml, ready-to-use)
11. Standard solution 1 (1 ml, ready-to-use) 0.0625 ng/ml AMOZ-NP
12. Standard solution 2 (1 ml, ready-to-use) 0.125 ng/ml AMOZ-NP
13. Standard solution 3 (1 ml, ready-to-use) 0.25 ng/ml AMOZ-NP
14. Standard solution 4 (1 ml, ready-to-use) 0.50 ng/ml AMOZ-NP
15. Standard solution 5 (1 ml, ready-to-use) 1.00 ng/ml AMOZ-NP
16. Standard solution 6 (1 ml, ready-to-use) 2.00 ng/ml AMOZ-NP

Storage

Store the kit at +2°C to +8°C in a dark place. For repeated use store kit components as specified under Reagent Preparation.
After the expiry date (see kit label) has passed, quality claims are not accepted.
Before opening the sealed plate, the plate should be at ambient temperature in order to avoid condensation in the ELISA.
Dilute the kit components immediately before use, but after the components are at ambient temperature.
The substrate chromogen solution can be stored in a refrigerator (+2°C to + 8°C) until the expiry date stated on the label.
Any direct action of light on the chromogen solution should be avoided.
Degeneration of the reagents may have occurred when the following phenomena are observed:
A blue colouring of the substrate solution before transferring it into the wells.
A weak or absent colour reaction of the zero standard (E450nm < 0.8).

Sensitivity

The LOD is determined under optimal conditions. Cut-off values need critical consideration.

General Description

The nitrofurans are a group of synthetic broad-spectrum antibiotics, which have been widely and effectively used for the prevention and treatment of gastrointestinal infections caused by Escherichia coli, Salmonella spp., Mycoplasma spp., Coccidia spp., coliforms and other protozoa in animal production and aquaculture. Moreover, nitrofurans have been employed as growth promoters in livestock. The four major nitrofurans are furazolidone, furaltadone, nitrofurantoin and nitrofurazone. They have been banned in the EU for use as veterinary drugs, due to their toxic an suspected carcinogenic and mutagenic properties (Commission Regulation 1442/95). In 2003 a definitive MRPL(Minimum Required Performance Limit) was set at 1 ng/g (ppb) in the EU for all four of the above mentioned nitrofurans in poultry and aquaculture products (Commission Decision 2003/181/EC). Various studies have demonstrated that the nitrofuran parent molecules are rapidly metabolised by animals and that their in vivo stability is not longer than a few hours. As a result, persistent protein-bound residues are formed. Unlike the parent molecules, these protein-bound metabolites are stable and persistent in the body. It is possible to free these residues from proteins by acid hydrolysis. Testing for the presence of nitrofurans is thus equivalent to testing for the presence of a part of the parent molecule, i.e. the free residue. Nitrofuran residues are found after administration of furaltadone (3-amino-5-morpholinomethyl-2-oxazolidinone = AMOZ), furazolidone (3-amino-2-oxazolidinone = AOZ), nitrofurantoin (1-aminohydantoin = AHD) and nitrofurazone (semicarbazide = SEM).

Standard Curve