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Fluorescence immunochromatographic assay method for quantitatively detecting heart fatty acid binding protein and kit for quantitatively detecting same

A technology of fluorescence immunochromatography and fatty acid combination, which is applied in the field of medical testing, can solve the problems of cumbersome operation steps, inaccurate quantification, and low sensitivity, and achieve the effects of expanding the detection range, improving sensitivity, and improving detection sensitivity

Active Publication Date: 2012-06-27
SHENZHEN KANGMEI BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Compared with the detection of fluorescence intensity, it has the disadvantages of low sensitivity and inaccurate quantification.
[0007] Therefore, based on the many advantages of quantum dots and immunochromatography, it is necessary to develop a method for the quantitative detection of early markers of myocardial injury (hFABP) using quantum dot fluorescence to make up for the pollution, cumbersome operation steps, high cost or sensitivity in the existing technology. The shortcomings of low and quantitative inaccuracy

Method used

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  • Fluorescence immunochromatographic assay method for quantitatively detecting heart fatty acid binding protein and kit for quantitatively detecting same
  • Fluorescence immunochromatographic assay method for quantitatively detecting heart fatty acid binding protein and kit for quantitatively detecting same
  • Fluorescence immunochromatographic assay method for quantitatively detecting heart fatty acid binding protein and kit for quantitatively detecting same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Example 1: Quantitative detection of hFABP by modifying antibodies to quantum dots by covalent cross-linking and using direct pre-wetting immunochromatography

[0069] (1) Modification of quantum dots and antibodies

[0070] Quantum dots with an emission wavelength of 650nm were mixed with 1mg / mL hFABP monoclonal antibody, and mixed with freshly prepared N-hydroxysuccinimide (NHS, 1mg / mL) and carbodiimide hydrochloride (EDC, 1mg / mL) under the action of room temperature for 4 to 5 hours, adding 1mol / L glycine to block, and separating and purifying with a chromatographic column or chromatographic column to obtain hFABP monoclonal antibody-modified quantum dots. Similarly, rabbit IgG-modified quantum dots were obtained. The fluorescence emission wavelength of the hFABP antibody-modified quantum dot is 650nm, and the fluorescence emission wavelength of the rabbit IgG-modified quantum dot is 570nm.

[0071] (2) Construction of the kit

[0072] Mix the two kinds of quantu...

Embodiment 2

[0084] Example 2: Quantitative detection of hFABP by modifying antibody to quantum dots with biotin-avidin system and using indirect pre-wetting immunochromatography

[0085] (1) Modification of quantum dots and antibodies

[0086] First, 1 mL of hFABP monoclonal antibody (1 mg / mL) was fully dialyzed with pH 9.0 sodium bicarbonate buffer, and 20-120 μl of N-hydroxysuccinimide biotin ester freshly prepared in dimethyl sulfoxide (DMSO) was added (NHSB, 1mg / mL), react at room temperature in the dark for 4h. Add 1mol / L NH 4 Cl, shaking reaction at room temperature for 10 min, then place the solution in a dialysis bag, dialysis and purification overnight, and concentrate with an ultrafiltration centrifuge tube to prepare the required concentration, and the obtained biotinylated hFABP monoclonal antibody.

[0087] Streptavidin-modified quantum dots with an emission wavelength of 900nm and biotinylated hFABP monoclonal antibody were mixed and reacted at a ratio of 1:3-1:12 for 30-6...

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Abstract

The invention discloses a fluorescence immunochromatographic assay method for quantitatively detecting hFABP (heart fatty acid binding protein) and a kit for quantitatively detecting the same. The fluorescence immunochromatographic assay method for quantitatively detecting the hFABP realizes quantitative fluorescence detection on the basis of optimizing components of a test strip by the aid of excellent fluorescent characteristics of quantum dots and by means of combining bicolor labeling technique and immunochromatographic assay. Compared with a conventional colloidal gold immunochromatographic assay method, the fluorescence immunochromatographic assay method has the advantages of fine labeling stability, low non-specificity, high sensitivity, wide linear range and accuracy in quantization. The kit is used for quantitatively detecting the hFABP, can be used for simultaneously detecting whole blood, blood serum and plasma samples, serves as a simple, accurate, specific and inexpensive detecting tool for early screening and prognosis evaluation of acute myocardial infarction, is applicable to hospitals at all levels, and is particularly beneficial to wide popularization in primary hospitals and clinics.

Description

technical field [0001] The invention relates to the field of medical testing, in particular to an early, sensitive and specific quantitative screening and detection of acute myocardial infarction by utilizing the immune response and fluorescence detection technology related to the human-shaped fatty acid binding protein. Background technique [0002] Acute coronary syndrome (ACS) is one of the most important acute events in coronary heart disease. It is mainly divided into acute myocardial infarction (AMI), unstable angina (UA) and sudden cardiac death (SCD). Both are higher. However, if high-risk patients can be identified early after onset and reperfusion therapy can be performed, the mortality and prognosis will be significantly improved. Among them, cardiac markers play a crucial role in the diagnosis and prognosis of ACS. Currently, commonly used cardiac markers include troponin (cTn), myoglobin (MYO) and creatine kinase isoenzyme (CK-MB). Because ACS has the charact...

Claims

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Application Information

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IPC IPC(8): G01N33/68
Inventor 王秀利
Owner SHENZHEN KANGMEI BIOTECH
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