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Fluorescence immunochromatographic assay and kit for quantitative detection of creatine kinase isoenzyme (CK-MB)

A technology of fluorescence immunochromatography and creatine kinase, which is applied in the field of medical testing, can solve the problems of expensive instruments, narrow linear range, and low sensitivity of creatine kinase isoenzyme detection, and achieve the effect of improving sensitivity and detection sensitivity

Active Publication Date: 2014-07-09
SHENZHEN KANGMEI BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] The technical problem to be solved in the present invention is to provide a fluorescent immunochromatographic method for quantitative detection of CK-MB in view of the shortcomings of low sensitivity, narrow linear range and expensive instruments in the prior art for detecting creatine kinase isoenzyme

Method used

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  • Fluorescence immunochromatographic assay and kit for quantitative detection of creatine kinase isoenzyme (CK-MB)
  • Fluorescence immunochromatographic assay and kit for quantitative detection of creatine kinase isoenzyme (CK-MB)
  • Fluorescence immunochromatographic assay and kit for quantitative detection of creatine kinase isoenzyme (CK-MB)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Example 1: Quantitative detection of CK-MB by modifying antibody to quantum dots by chemical cross-linking method and using direct pre-wetting immunochromatography

[0070] (1) Modification of quantum dots and antibodies

[0071] Using human CK-MM and CK-BB as antigens, goat anti-human CK-MM antibodies and CK-BB antibodies can be obtained by immunization. Since CK-MB has two subunits, M and B, the CK-MM antibody and CK-BB antibody can bind to the M and B subunits of CK-MB respectively to form a double-antibody sandwich structure. ,

[0072] Add 60pmol quantum dots with emission wavelength of 650nm, 10μg EDC and 15μg NHS solution and 10-30μg goat anti-human CK-BB antibody solution to pH 7.4 phosphate buffer solution, mix well and react at room temperature for 4h, add 1mg glycine to block. Separating and purifying with a chromatographic column or a chromatographic column to obtain the quantum dot modified by the CK-BB antibody. Similarly, quantum dots modified with goa...

Embodiment 2

[0082] Example 2: Quantitative detection of CK-MB by modifying antibody to quantum dots by biotin-avidin method and using indirect pre-wetting immunochromatography

[0083] (1) Synthesis and modification of quantum dots

[0084] According to the method of Example 1, streptavidin-modified quantum dots were prepared. Similarly, quantum dots modified with goat anti-rabbit antibody were obtained.

[0085] Dilute CK-BB monoclonal antibody to 1mg / ml with 0.1mol / L sodium bicarbonate buffer (pH 8.0), dissolve 1mg N-hydroxysuccinimide biotin (NHSB) with 1ml dimethyl sulfoxide (DMSO), Take 1ml monoclonal antibody solution and add 20-120μl NHSB solution, react at room temperature for 4 hours, add 1mol / L NH 4 Cl, stirred at room temperature for 10 min. Purify by ultrafiltration centrifugation to remove free biotin to obtain biotinylated mAb.

[0086] Streptavidin-modified quantum dots and biotinylated monoclonal antibodies are mixed at a ratio of 1:3 to 1:12 to obtain CK-BB monoclonal...

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Abstract

The invention discloses a fluorescence immunochromatographic assay and kit for quantitative detection of acute myocardial infarction marker-creatine kinase isoenzyme (CK-MB). The fluorescence immunochromatographic assay for quantitative detection of the CK-MB realizes fluorescent quantitative detection by utilizing excellent fluorescent properties of quantum dots and combining a bicolour marking technology and an immunochromatographic assay on the basis of optimizing each component of a test strip. Compared with the common collaurum immunochromatographic assay, the fluorescence immunochromatographic assay has the advantages of good mark stability, low nonspecificity, high sensitivity, wide linear range and accuracy in quantification. The kit disclosed by the invention is used for carrying out quantification detection on the CK-MB, is suitable for detection of whole blood, blood serum and plasma samples, can provide a reference for cardiovascular and cerebrovascular disease diagnosis, and is widely applied to primary hospitals and clinics.

Description

technical field [0001] The invention relates to the field of medical testing, in particular to a fluorescent immunochromatographic method for quantitatively detecting acute myocardial infarction marker creatine kinase isoenzyme and a kit thereof. Background technique [0002] At present, cardiovascular disease has become the largest potential killer of adults worldwide. In the United States alone, more than 70 million people suffer from heart disease, and about 6 million people go to the emergency room due to chest pain every year. Acute myocardial infarction ( The death toll of AMI) has exceeded 500,000. In my country, the prevalence, incidence and risk factors of cardiovascular diseases are on the rise. According to statistics in recent years, the mortality rate of cardiovascular disease accounts for 40% of the population deaths, which is higher than that of European and American countries, and belongs to the country with high incidence of cardiovascular disease. Therefore...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/573
Inventor 王东
Owner SHENZHEN KANGMEI BIOTECH
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