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Colloidal gold test strip for semi-quantitatively detecting concentration of tacrolimus drug in human whole blood and detection method

A semi-quantitative detection and tacrolimus technology, applied in the field of tacrolimus drug detection, can solve the problems of semi-quantitative detection and semi-quantitative detection, and achieve high specificity, high sensitivity, fast and easy operation Effect

Active Publication Date: 2010-10-06
同昕生物技术(北京)有限公司
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  • Application Information

AI Technical Summary

Problems solved by technology

[0008] So far, there is no reported method for the efficient and convenient semi-quantitative detection of tacrolimus drug concentration in whole blood, and there is no method for semi-quantitative detection of tacrolimus drug concentration in whole blood combined with competitive immunobinding technology and colloidal gold detection technology. Reports on Quantitative Detection

Method used

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  • Colloidal gold test strip for semi-quantitatively detecting concentration of tacrolimus drug in human whole blood and detection method
  • Colloidal gold test strip for semi-quantitatively detecting concentration of tacrolimus drug in human whole blood and detection method
  • Colloidal gold test strip for semi-quantitatively detecting concentration of tacrolimus drug in human whole blood and detection method

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preparation experiment example 1

[0048] Step 1. Tacrolimus Activation and Carrier Protein Coupling

[0049] In the present invention, the coupling process of tacrolimus hapten and carrier protein refers to the method in US Patent No. 5,164,495. Simply speaking, in the presence of triethylamine, tacrolimus molecules are dry mixed with succinic acid, and a catalyst is added. 4-Dimethylaminopyridine, purified after reaction to obtain tacrolimus half ester, then activated, and then reacted with carrier protein to obtain a conjugate.

[0050] The tacrolimus antigen of the present invention is a conjugate of a tacrolimus hapten and a carrier protein molecule, and the carrier unit includes bovine serum albumin, human serum albumin, ovalbumin and other common carrier proteins.

[0051] The specific operation is as follows:

[0052] Weigh 250mg FK506, 15mg succinic anhydride, 6mg pyridine (4-dimethylaminopyridine) and dissolve in 6.25ml freshly distilled dichloromethane. Add 42ul triethylamine while stirring. Stirr...

experiment example 2

[0067] Experimental example 2 detection sample process

[0068] 1.1. Sample processing

[0069] Take 0.5ml of fingertip blood with a blood collection needle and a quantitative pipette, quickly add it to the centrifuge tube containing the sample treatment solution, and shake it vigorously for 5 minutes to rupture the red blood cells and release tacrolimus.

[0070] 1.2. Detection with test strips

[0071] Use a quantitative plastic tube to take 120ul of the treated sample and drop it into the sample hole of the test strip, start timing, wait for the red band to appear, and read the value within 10 minutes. If it exceeds 20 minutes, the result will be invalid.

[0072] 1.3. Interpretation of results

[0073] This reagent adopts the competition method, and the three concentration gradient marking lines designed are 5ng / ml, 10ng / ml, and 20ng / ml, and the results are interpreted as follows:

[0074] When there are four lines containing quality control lines, such as Figure 4a ,...

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Abstract

The invention relates to a colloidal gold test strip for semi-quantitatively detecting the concentration of tacrolimus drug in human whole blood and a reagent kit, belonging to the field of medical detection. A membrane reacting system of the colloidal test strip comprises a colloidal gold combining pad and a detection pad. The colloidal gold test strip is characterized in that the colloid gold combining pad is coated by anti-tacrolimus antibody marked by colloid gold; and the detection pad is provided with a detection line, and tacrolimus-carrier protein conjugate is fixed on the detection line. The detection method does not need steps of precipitating organic solvent, centrifugation and the like during detection, and can semi-quantitatively detect tacrolimus in whole blood simply, rapidly, accurately and effectively.

Description

technical field [0001] The invention relates to the detection of tacrolimus drug in human whole blood, in particular to a colloidal gold test strip for semi-quantitative detection of tacrolimus drug concentration in human whole blood. Background technique [0002] Tacrolimus is a macrolide immunosuppressant that was first used clinically in 1989. Tacrolimus binds to its binding protein FKBP12, and accumulates in cells to produce effect. Complexes such as FKBP12-tacrolimus can specifically bind and inhibit the activity of calcineurin, block the activation process of T cells, and also have T-cell dependent antibodies that tacrolimus does not possess It produces and blocks the transcription of cytokines, presents a double immunosuppressive effect, and its immunosuppressive effect is 100 times stronger than that of tacrolimus. It is mainly used clinically for the suppression of immune rejection after liver and kidney transplantation. The individual absorption and clearance rat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/577G01N1/28
Inventor 李全焦守恕
Owner 同昕生物技术(北京)有限公司
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