1. Principle
This test kit is based on the competitive enzyme immunoassay for the detection of Gentamicin in the sample. The coupling antigen is pre-coated on the micro-well stripes. The Gentamicin in the sample and pre-coated coupling antigen on the micro-well stripes compete for the anti-Gentamicin antibody. After the addition of the enzyme conjugate, the TMB substrate is added for coloration. The optical density (OD) value of the sample has a negative correlation with the Gentamicin in it. The value is compared to the standard curve and the Gentamicin concentration is subsequently obtained.
2. Technical specifications
Sensitivity: 0.1 ppb
Detection limit
Chicken............................................................................................ 4 ppb
Liver................................................................................................. 4 ppb
Recovery rate
Chicken......................................................................................... 70±10%
Liver.............................................................................................. 70±10%
Cross-reaction rate
Gentamicin........................................................................................ 100%
Streptomycin...................................................................................... <1%
Dihydrostreptomycin........................................................................... <1%
Neomycin........................................................................................... <1%
3. Components
1) Micro-well strips: 12 strips with 8 removable wells each
2) 6× standard solution (1 mL each): 0 ppb, 0.1 ppb, 0.3 ppb, 0.9 ppb, 2.7 ppb, 8.1 ppb
3) Enzyme conjugate (7 mL)............................................................... red cap
4) Antibody working solution (7 mL).................................................. blue cap
5) Substrate A solution (7 mL).......................................................... white cap
6) Substrate B solution (7 mL)......................................................... black cap
7) Stop solution (7 mL).................................................................. yellow cap
8) 20× concentrated washing buffer (40 mL)..................................... white cap
9) 2× concentrated redissolving solution (50 mL)..................... transparent cap
4. Materials required but not provided
1) Equipments: microplate reader (450 nm / 630 nm), votex, centrifuge, homogenizer, measuring pipets and balance (a sensibility reciprocal of 0.01 g)
2) Micropipettors: single-channel 20-200 µL and 100-1000 µL, and multi-channel 250 µL;
3) Reagents: Na2HPO4·12H2O, NaH2PO4·2H2O
5. Sample pre-treatment
Instructions
The following points must be dealt with before the pre-treatment of any kind of sample:
1) Only the disposable tips can be used for the experiments and the tips must be changed when used for absorbing different reagents;
2) Before the experiment, each experimental utensil must be checked to be clean and should be re-cleaned if necessary, in order to avoid the contamination that interferes with the experimental results.