A kit for quantitative determination of interleukin-6 and its preparation method
A technology for quantitative determination of interleukin, applied in the field of quantitative detection of interleukin 6 in blood, can solve the problems of unfavorable high-throughput automatic detection, poor specificity, low sensitivity, etc., and achieve optimized chemiluminescence enhancement system, small variation, and high sensitivity Effect
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Embodiment 1
[0049] Embodiment 1: the configuration of various buffers, specifically as follows:
[0050] 1. Tris salt buffer
[0051] Add 12.12g of Tris and 5.82g of sodium chloride into 1L of purified water, stir well until completely dissolved, and adjust the final pH to 7.5 with hydrochloric acid.
[0052] 2. Preparation of calibrator buffer
[0053] Add 0.01g of tetracycline and 0.1g of neomycin sulfate to 1L of newborn bovine serum, fully dissolve and process through 0.22μm filter membrane to prepare.
[0054] 3. Anti-reagent buffer
[0055] Add 30.5mg of Tris, 0.03g of tetracycline, 3g of sheep serum, 8g of newborn bovine serum, and 3g of horse serum into 1L of purified water, stir well until completely dissolved;
[0056] 4. Magnetic particle buffer
[0057] Add 12.12mg of Tris, 5.82mg of sodium chloride, and 50g of methyl cellulose ether into 1L of purified water, stir well until completely dissolved.
[0058] 5. Luminescence substrate buffer
[0059] Add 50.21g of Tris, 5.8...
Embodiment 2
[0062] Example 2: Preparation of Interleukin-6 Quantitative Assay Kit
[0063] 1. Preparation of calibrators and quality controls
[0064] Firstly, the interleukin-6 was dissolved in the standard buffer solution, and prepared into the calibration product and the quality control product with the target concentration as shown in Table 1; the interleukin-6 used in this example was purchased from the manufacturer fitzgerald company.
[0065] Table 1 Preparation of Calibrators and Quality Controls
[0066]
[0067] 2. The preparation method of the anti-reagent is as follows:
[0068] 1) Coupling of fluorescein isothiocyanate and interleukin-6 antibody to obtain fluorescein isothiocyanate-labeled interleukin-6 coated antibody:
[0069] Firstly, fluorescein isothiocyanate solution was prepared into a concentration of 2.5mg / mL fluorescein isothiocyanate solution with anti-reagent buffer, and then according to the mass ratio of interleukin 6: fluorescein isothiocyanate solution=1:...
Embodiment 3
[0080] Embodiment 3: Utilize the method for detecting interleukin-6 with the interleukin-6 quantitative assay kit described in embodiment 2, the method comprises the steps:
[0081] 1) Take three test tubes and add 15 μL interleukin-6 calibrator, 15 μL interleukin-6 quality control substance, and 15 μL sample to be tested;
[0082] 2) Add 60 μL of anti-reagent to each test tube, cover the test tube with a plastic film, shake the test tube gently for 30 seconds, and place it in a water bath at 37°C for 5 minutes;
[0083] 3) Add 30 μL of magnetic particle reagent to each test tube, cover the test tube with a plastic film, shake the test tube gently for 30 seconds, and place it in a water bath at 37°C for 5 minutes;
[0084] 4) Precipitate the test tube on the magnetic separator for 2 minutes, slowly invert the test tube and the magnetic separator, pour out the supernatant, put the inverted test tube together with the magnetic separator on the filter paper, and pat the bottom of...
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