Enrofloxacin Rapid test strip

1.Principle

The Enrofloxacin rapid test strip is based on immuno-chromatographic technology, Enrofloxacin in sample in the process of flow combine with the colloidal gold-labeled monoclonal antibody, inhibits the antibody combining with Enrofloxacin - BSA conjugate binding on the NC membrane test line. If the content of Enrofloxacin in the sample is high, T line is lighter than C line or T line is invisible, the result is positive. On the contrary, T line is same as C line or is darker than C line, the result is negative.

The detection limit of Enrofloxacin is 10ppb, (There is cross-reaction with Ciprofloxacin, danofloxacin, Flumequine etc.)

2.Components

3.Sample collecting and preparation

Collect the fish, shrimp and crab etc. into clean container or sealed bag; If can’t test on time, the sample should remove fat, use scissors to shred or use mincer to mince, store at below -20℃, avoid repeated freezing and thawing.

3.1 Tissue (fish, chicken, pig)

(1) Take certain quantity tissue (fish, chicken, pig), remove fat, use scissors to shred or use mincer to mince;

(2) Take 5g sample into 50ml centrifuge tube;

(3) Add 10ml 0.2mol (PH=11.4) NaOH (It can use HCl to adjust PH value), shake to mix evenly for 2min;

(4) Centrifuge at 4000rpm fro 10min, take 7ml supernatant into another 50ml centrifuge tube, and add 6ml CH2Cl2, mix up-and-down for 10s (Avoid shake violently);

(5) Centrifuge at 4000rpm for 5 min, take 3ml down-layer solution into 5ml centrifuge tube, blow to dry by nitrogen or air at 60℃;

(6) Use Micropipettes to add 500ul Enrofloxacin PBST buffer, then 0.5ml Hexane to redissolve, mix up-and-down for 1 min;

(7) Centrifuge at 4000rpm for 1 min (If stratification is not clear, extend to 5min) or be static until obvious stratification, take 150ul down-layer solution to test.

3.2 Shrimp, crab

(1) Take certain quantity aquatic products (shrimp, crab), remove fat, use scissors to shred or use mincer to mince;

(2) Take 4g sample into 50ml centrifuge tube;

(3) Add 8ml CH2Cl2, shake to even for 2min;

(4) Centrifuge at 4000rpm for 5 min, take 4ml down-layer solution into 5ml centrifuge tube, blow to dry by nitrogen or air at 60℃;

(5) Use Micropipettes to add 500ul Enrofloxacin PBST buffer, then 0.5ml Hexane to redissolve, mix up-and-down for 1 min;

(6) Centrifuge at 4000rpm for 1 min, take 150ul down-layer solution to test.

4.Operation procedures

1) Return test strip and reagents to room temperature before testing;

2) Take out the test strip from the original package, use it in half hour;

3) Put test card on desk flatly, Absorb 150μL ready-to test solution into gold label well, use small dropper to blow and beat until the red material is completely dissloved, wait to react for 2min,

4) Use dropper absorb all sample from the well, add it into sample collecting well (as following picture), timing after adding sample.

5) Read the result in 8-10 minutes, it is invalid after 15min.

Note: S means Sample Colleting Region

     C means Control Line

     T means Test Line

5.Test Result Interpretation

5.1 Negative (—):  T line is darker or same as C line, it means the Enrofloxacin concentration is lower than the detection limit.

5.2 Positive (+): T line is lighter than C line or T line is invisible, it means the Enrofloxacin concentration is higher than the detection limit.

5.3 Invalid: C line is invisible, maybe it is the operation error or test strip is invalid. Read the manuals, use new test strip to test again.

6.Precautions

1) Use the test strip at room temperature for once, do not repeat use.

2) Do not touch the white membrane surface in the middle of test card, avoid sunshine directly and fan blow directly.

3) For positive result, use this test card test again.

7.Storage and expiry date

Storage:  Store at 4-30 ℃ in dark, no frozen. 

Expiry date: 12 months; date of production is on box.