Magnetic bead time resolution fluorescence immunoassay quantitative determination CK-MB (creatine kinase-MB) kit

A time-resolved fluorescence and immunoquantitative technology, applied in measurement devices, analytical materials, instruments, etc., can solve the problems of narrow quantitative measurement range of labeled enzymes and substrates, unsuitable for use in primary hospitals, poor sensitivity and accuracy, etc. Long fluorescence decay time, no radioactive pollution, and the effect of reducing background

Inactive Publication Date: 2018-07-27
GUANGZHOU BIOKEY HEALTH TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Immunosuppressive method is easily interfered by CM-BB and giant CK, which limits the application of this method
Enzyme-linked immunoassay has the characteristics of long validity period of reagents, good specificity, and the results can be measured by instruments. However, due to the relatively low sensitivity, the shortcomings of the labeled enzyme and substrate that can be quantitatively determined and the narrow range of instrument determination are limited. Its application in microimmunoquantitative assay
Although the immunochromatography method is simple to operate, its sensitivity and accuracy are poor
Although chemiluminescence immunoassay has high sensitivity, it takes a long time to detect and the equipment is expensive, so it is not suitable for primary hospitals

Method used

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  • Magnetic bead time resolution fluorescence immunoassay quantitative determination CK-MB (creatine kinase-MB) kit
  • Magnetic bead time resolution fluorescence immunoassay quantitative determination CK-MB (creatine kinase-MB) kit
  • Magnetic bead time resolution fluorescence immunoassay quantitative determination CK-MB (creatine kinase-MB) kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] In this embodiment, the magnetic bead time-resolved fluorescence immunoquantitative detection kit for CK-MB consists of a reagent strip and a CK-MB calibrator.

[0028] Wherein, the shape of the reagent strip is fan-shaped, and 1 to 13 holes are arranged sequentially from left to right. Wells 1 and 2 are test wells for storing immunomagnetic beads and reagent wells; wells 3 and 4 are fluorescently labeled wells for storing europium-labeled antibodies; wells 6 and 7 are cleaning solution wells, It is used to store cleaning solution; No. 8 and No. 9 are sample diluent holes, which are used to store sample diluent; No. 12 is an enhancement fluid hole, used to store enhancement fluid; No. 5, 10, 11, and 13 are preparatory holes. A magnet can be stored between the 1st and 2nd wells for magnetic separation experiments. The 1st and 2nd wells can store a liquid volume of 300 μl; the 3rd and 4th wells can be disassembled from the entire reagent strip to become independent compon...

Embodiment 2

[0060] The making and detection of embodiment 2 reagent strips

[0061] The semi-finished reagent strips in this example are assembled through the following procedures: 50 μL CK-MB immunomagnetic beads, 200 μL europium-labeled CK-MB antibody, 2000 μL cleaning solution, 2000 μL Cleaning solution, 400 μL enhancement solution, and then seal with a film sealing machine. The sealing film is coated with product information that can be scanned and identified by an automatic fluorescence detection analyzer, including enterprise standard curve, batch, production date, and expiration date. The finished reagent strips, packaged CK-MB calibrator and other accessories are assembled into a kit.

[0062] Sample testing:

[0063] ①Add sample:

[0064] Put the sample or calibrator to be tested into the loading system of the automatic instrument, insert the reagent strip into the reagent strip slot, and the instrument will automatically identify the product information of the sealing film. A...

Embodiment 3

[0075] The comparison of embodiment 3 and commercially available kit

[0076] The commercially available creatine kinase assay kit (chemiluminescence microparticle immunoassay) (Abbott Trading Co., Ltd.) and the method in Example 1 were used to detect the sample (the concentration range of CK-MB in the sample was 1-300 ng / mL) Repeat three times to verify the correctness of the detection results of the kit of the present invention, the results are shown in Table 1 below.

[0077] Table 1 Sample test results

[0078]

[0079] Compared with commercially available reagents, the deviation of this reagent is less than ±5%, and the detection result of this reagent is accurate and reliable.

[0080] The commercially available creatine kinase assay kit (chemiluminescent microparticle immunoassay) (Abbott Trading Co., Ltd.) and the reagents in Example 1 were used to detect 200 clinical samples, and the test results are shown in Table 2 below.

[0081] Table 2 Test results of clinic...

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Abstract

The invention discloses a magnetic bead time resolution fluorescence immunoassay quantitative determination CK-MB kit. The CK-MB kit comprises an immunomagnetic bead coating a CK-MB monoclonal antibody, a CK-MB standardized product solution, a europium-marked CK-MB monoclonal antibody solution, washing liquid and enhancement liquid. The immunomagnetic bead coating the CK-MB monoclonal antibody isa covalent conjugate of a superpara magnetic bead modified by a functional group and with the diameter being 1-3 microns and the CK-MB monoclonal antibody. The kit has the high sensibility, the sensibility of CK-MB is 1ng / mL, and a blood serum (plasma) does not need to be diluted; the determination time is short, and a report can be resulted within 30 minutes; the demanding amount of the sample isless, and only 50 microliters are needed for one-time sample loading; and the kit is equipped with a full-automatic time resolution immune analysis meter, operation is easy, no artificial error exists, and labor is saved. The kit reasonably utilizes the space of a reagent strip, the structure of the reagent strip is more compact, the reagent strip can be transported more easily, and used conveniently, the operation is simple, and the stability is good.

Description

technical field [0001] The invention relates to the technical field of in vitro reagent detection, in particular to a magnetic bead time-resolved fluorescence immunoquantitative detection kit for CK-MB. Background technique [0002] Creatine Kinase (CK) is usually found in the cytoplasm and mitochondria of animal heart, muscle, brain and other tissues. It is an important energy-regulating enzyme that is directly related to intracellular energy operation, muscle contraction, and ATP regeneration. It reversibly catalyzes the transphosphorylation reaction between creatine and ATP. CK consists of two subunits, M and B, and is divided into three basic isozymes: muscle type (CK-MM), brain type (CK-BB), and hybrid type (CK-MB). [0003] Human myocardial tissue contains a large number of enzymes, including creatine kinase (CK), creatine kinase isoenzyme (CK-MB), lactate dehydrogenase (LDH), etc. Under the influence of pathological changes, different myocardial enzymes will Obvious...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577
CPCG01N33/577
Inventor 李根平解巧丽
Owner GUANGZHOU BIOKEY HEALTH TECH CO LTD
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