Human myeloperoxidase enzyme activity and total quantity detection kit, detection method and preparation method
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A myeloperoxidase and detection kit technology, which can be applied to measurement devices, biological material analysis, instruments, etc., can solve irrelevant problems and achieve the effects of convenient operation, improved specificity and accuracy, and shortened measurement time.
Active Publication Date: 2017-06-30
海格德生物科技(深圳)有限公司
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[0006] Based on this, it is necessary to provide a new detection method for human myeloperoxidase to solve the problem that MPO content is not related to MPO activity, and remove the interference of other impurities in the sample, greatly improve the specificity and accuracy, shorten the Measurement time, reduced kit cost, and easy handling
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example 1
[0103] Mix anti-MPO non-inhibitory antibody (10ug / ml) with 50mM (pH9.6) carbonate buffer well, add 100ul to each well of the microplate, 4°C, overnight;
[0104] Wash once with washing solution, add 200 μl blocking solution to each well, and block for 2 hours at 4°C;
[0105] Dry the blocking solution and wash once;
[0106] Add 80ul sample diluent and 20ul serum or plasma sample to the well mixture, and react for 30 minutes at room temperature;
[0107] Shake off the liquid and wash twice with washing liquid;
[0108] Add 200ul buffer solution (sodium citrate buffer, 10mmol / L phenol), add 50ul enzyme decomposition substrate (H2O220mmol / L), mix, and place at room temperature for 1.5min, 2min, 3min respectively
[0109] Measure the OD value with a wavelength of 505nm;
[0110] Calculation of MPO enzyme activity test results: take the antigen concentration of the calibrator (0, 50, 100, 250, 500, 1000ng / ml) as the abscissa, and the corresponding OD value of the calibrator as ...
example 1
[0125] Including the preparation of MPO solid-phase carrier, the preparation of MPO enzyme-labeled antibody reagent, the preparation of MPO calibrator, and the preparation of other components (including chromogenic substrate A, chromogenic substrate B, stop solution, concentrated washing solution (20X)) , the main process is as follows:
[0126] 1. Preparation of MPO solid phase carrier
[0127] 1. Preparation of solid-phase carrier coating solution and blocking solution: see Table 1 and Table 2 below for the formula.
[0128] Table 1. Coating solution formula
[0129] Raw material name Prepare 1000ml dosage Na 2 CO 3
1.59g NaHCO 3
2.93g
[0130] First add 900ml of purified water, after all the substances are dissolved, adjust the pH value to 9.6±0.1, and then adjust the volume to 1000ml.
[0131] Table 2. Blocking solution formula
[0132] Raw material name Prepare 1000ml dosage BSA 10.0g sucrose 52.0g cas...
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Abstract
The invention discloses a human myeloperoxidase enzyme activity and total quantity detection kit, a detection method and a preparation method. The detection kit comprises a solid-phase carrier, a zymolysis substrate, buffer solution, an enzyme antibody reagent, a plurality of calibration products, cleaning solution, a chromogenic substrate and stop solution, the solid-phase carrier is coated with 200-400ng / hole of anti-MPO (myeloperoxidase) non-restraining antibodies, the zymolysis substrate comprises 0.1-50mmol / L of H2O2, the enzyme antibody reagent contains anti-MPO (myeloperoxidase) restraining antibodies marked by horse radish peroxidase, the usage amount of the antibodies is 1:2000-1:8000, the antigen content of myeloperoxidase of the calibration products is 0-1000ng / ml, the chromogenic substrate at least comprises chromogenic substrate A solution and chromogenic substrate B solution, the chromogenic substrate A solution contains 0.015-0.02% of peroxide, and the chromogenic substrate B solution contains 0.25-0.32g / L of TMB (tetramethylbenzidine). According to the detection kit, the conditions of the myeloperoxidase in human bodies can be accurately responded, and the detection kit provides more accurate basis for clinical diagnosis and early warning of coronary arteries diseases, particularly, and unstable coronary heart diseases.
Description
technical field [0001] The invention relates to the field of in vitro diagnosis, in particular to a human myeloperoxidase activity and total amount detection kit, a corresponding human myeloperoxidase activity and total amount detection method, and a preparation method. Background technique [0002] Human myeloperoxidase (Myeloperoxidase, MPO), also known as peroxidase, is an important iron-containing lysosome, present in the azurophilic blue cells of myeloid cells (mainly neutrophils and monocytes). In the granule, it is a specific marker of myeloid cells and is an important part of the human immune system. When some inflammation occurs, MPO is released by activated neutrophils and monocytes, which can convert hydrogen peroxide and chloride ions into hypochlorous acid (HClO), a strong oxidant that can kill microorganisms, to kill Kill pathogens. [0003] Medical research at home and abroad shows that the increase of MPO concentration in blood is closely related to the for...
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