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Creatine kinase isoenzyme detection kit

A detection kit, creatine kinase technology, applied in biological tests, measuring devices, material inspection products, etc.

Active Publication Date: 2018-09-18
BEIJING STRONG BIOTECH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, most of the detection reagents used in clinical practice have not taken measures to prevent RF interference (Research progress on the interference of rheumatoid factors on immunoassays, Medical Review 2015 Vol 21, 19: 3495)

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Embodiment 1. Preparation of the first reagent

[0064] 1. Weigh 17.87g HEPES, 13.15g NaCl, 9g PEG20000, 15g Tween 80, 1.5g sodium azide, 7.5g BSA, 15mL blocker (5mg / ml), dissolve in 1.0L double distilled water, adjust the pH to 7.2, set the volume to 1.5L to get the first reagent (1):

[0065]

[0066] 2. Alternatively, weigh 9.09g Tris, 13.15g NaCl, 9g PEG20000, 15g Tween80, 1.5g sodium azide, 7.5g BSA, 10mL blocker (same as above), dissolve in 1.0L double distilled water, adjust the pH to 7.2 , dilute to 1.5L to prepare the first reagent (2):

[0067]

[0068] 3. Or, weigh 5.63g glycine, 13.15g NaCl, 9g PEG20000, 15g Tween 80, 1.5g sodium azide, 7.5g BSA, 10mL blocker (same as above), dissolve in 1.0L double distilled water, adjust the pH to 7.2, dilute to 1.5L to prepare the first reagent (3):

[0069]

Embodiment 2

[0070] Example 2. Preparation of antibody-coated particles

[0071] 1. Dilute the particles (300nm, carboxyl) to 1% (w / v) with HEPES buffer;

[0072] 2. Dilute the antibody (mouse monoclonal antibody) to 0.5mg / ml with HEPES buffer;

[0073] 3. Add EDAC aqueous solution with a concentration of 0.1% to 1% (w / v) to the granules in step 1, and react in a constant temperature shaker at 37°C for 30min;

[0074] 4. After the reaction is over, add the antibody in step 2 and react in a constant temperature shaker at 37°C for 3 hours;

[0075] 5. Then add the sealing agent and leave it overnight at room temperature;

[0076] 6. Add the cleaning solution to wash and centrifuge the particles sealed overnight for 3 times, save them for later use.

[0077] The order of steps 1 and 2 is interchangeable.

Embodiment 3

[0078] Embodiment 3. Preparation of the second reagent

[0079] 1. Add 400 mL of double distilled water to the granules prepared in Example 2, add 18.76 g of glycine, 2.5 g of BSA, 0.5 g of sodium azide, and 50 g of sucrose, stir and mix well, adjust the pH to 7.5, and add double distilled water to 500 mL , sonication until the absorbance at the dominant wavelength does not change substantially to obtain the second reagent (1):

[0080]

[0081] 2. Alternatively, add 400 mL of double distilled water to the granules prepared in Example 2, add 5.96 g of HEPES, 2.5 g of BSA, 0.5 g of sodium azide, and 50 g of sucrose, stir and mix, adjust the pH to 7.5, and add double distilled water to 500mL, sonicate until the absorbance at the main wavelength does not change substantially to obtain the second reagent (2):

[0082]

[0083] 3. Alternatively, add 400mL double distilled water to the granules prepared in Example 2, add 3.03g Tris, 2.5g BSA, 0.5g sodium azide, 50g sucrose, s...

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Abstract

The invention relates to a creatine kinase isoenzyme detection kit. The kit includes a first reagent and a second reagent. The first reagent comprises a buffer solution, a preservative, a coagulant, asurfactant, a protective agent and a blocker; and the second reagent comprises the buffer solution, a stabilizer, the preservative, the protective agent, polystyrene latex particles and a creatine kinase isozyme antibody. The kit has the advantages of good stability and strong specificity, can be applied to biochemical analyzers in order to carry out large-batch detection, and can replace chemiluminescence products to reduce the detection cost in hospitals.

Description

technical field [0001] The application belongs to the field of in vitro diagnostic reagents, and specifically relates to the detection of creatine kinase isoenzyme by immunoturbidimetric method. Background technique [0002] CK was first studied in 1960 and thought it could be used to diagnose ACS, but it was replaced by creatine kinase isoenzyme (CKMB) after 1970 due to its poor specificity. Subsequently, CKMB was recommended as the gold standard for the diagnosis of myocardial infarction (AMI) due to its good specificity and sensitivity. [0003] However, since small amounts of CKMB are also present in skeletal muscle, CKMB will be higher in skeletal muscle in children. Therefore, the specificity is not good, and there is a risk of misdiagnosis. At present, it has been replaced by the new gold standard marker cTnI. [0004] Nevertheless, CKMB still has important clinical significance in the diagnosis of ACS and AMI. First, when myocardial injury triggers chest pain, CK...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/573
CPCG01N33/573G01N33/6854G01N33/54313C12Q1/50G01N33/54306G01N2333/9123
Inventor 熊争平果玮刘瑶刘希
Owner BEIJING STRONG BIOTECH INC
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