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.