Chemiluminiscence detection kit for tacrolimus and preparation method of chemiluminiscence detection kit
A technique for chemiluminescence detection and tacrolimus, which is applied to the chemiluminescence detection kit of tacrolimus and the field of preparation thereof, and can solve the problem of slow time-resolved fluorescence immunoassay, low sensitivity of enzyme-linked immunoassay, and radioactivity. The immunological method has many problems such as many operating steps, and achieves the effect of fast and convenient detection, stable luminescence value, and improved sensitivity.
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Embodiment 1
[0025] Embodiment 1: The formation and preparation of kit 1
[0026] 1. Kit 1 set-up
[0027] Construct a chemiluminescence detection kit for tacrolimus so that it contains the following components:
[0028] Tacrolimus antigen coupled with carboxyl magnetic beads;
[0029] Acridinate-labeled tacrolimus antibody;
[0030] Chemiluminescence pre-excitation solution A and chemiluminescence excitation solution B;
[0031] Tacrolimus series calibrator solution;
[0032] Washing fluid.
[0033] 2. Preparation of Magnetic Microparticle Suspension Conjugated with Tacrolimus Antigen
[0034] (1) Take 1 mg of carboxyl magnetic particles in a 0.5 mL centrifuge tube, add 200 μL of 0.1 mol / L MES (pH 5.0) buffer, vortex and mix, place on a magnetic stand, and let the magnetic particles stand for 5 min to make the magnetic particles Separate from the liquid, discard the supernatant, wash 3 times, add 200 μL of MES buffer (pH 5.0), and vortex.
[0035] (2) Add 20 μg of tacrolimus antige...
Embodiment 2
[0052] Embodiment 2: The formation and preparation of kit 2
[0053] 1. Assembly of Kit 2
[0054] Construct a chemiluminescence detection kit for tacrolimus so that it contains the following components:
[0055] Carboxyl magnetic beads coupled with tacrolimus antibody;
[0056] acridinium ester-labeled tacrolimus antigen;
[0057] Tacrolimus series calibrator solution;
[0058] Chemiluminescence pre-excitation solution A and chemiluminescence excitation solution B;
[0059] Washing fluid.
[0060] 2. Preparation of Magnetic Microparticle Suspension Conjugated with Tacrolimus Antibody
[0061] (1) Take 1 mg of carboxyl magnetic particles in a 0.5 mL centrifuge tube, add 200 μL of 0.1 mol / L MES (pH 6.0) buffer, vortex and mix, place on a magnetic stand, and let the magnetic particles stand for 5 min to make the magnetic particles Separate from the liquid, discard the supernatant, wash 3 times, add 200 μL of MES (pH 6.0) buffer, and vortex.
[0062] (2) Add 15 μg of tacro...
Embodiment 3
[0077] Example 3: Detection of Tacrolimus in Actual Samples
[0078] The operating procedures for the use of the tacrolimus quantitative detection kit of the present invention are as follows:
[0079] Detection of the kit
[0080] (1) Add 20 μL of the sample to be tested, 150 μL of the coupled magnetic particle suspension, and 150 μL of the acridinium ester marker into the reaction tube in sequence, shake and mix well, and incubate at 37°C for 15 min.
[0081] (2) Separate and wash 5 times.
[0082] (3) Fully shake the washed reaction container to disperse the magnetic particles.
[0083] (4) Add 100 μL of chemiluminescence pre-excitation solution A, add 100 μL of chemiluminescence excitation solution B after 1 s, and measure the relative luminescence intensity. The content of tacrolimus in the sample is proportional to the relative luminescence intensity.
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