Fluorescence immunochromatographic assay and kit for quantitatively detecting cardiac troponin T

A technology of fluorescence immunochromatography and cardiac troponin, which is applied in the field of medical testing, can solve the problems of low sensitivity and inaccurate quantification of cardiac troponin T, and achieve the effects of expanding the detection range, improving sensitivity, and improving quantitative accuracy

Active Publication Date: 2014-05-07
SHENZHEN KANGMEI BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] The technical problem to be solved in the present invention is to provide a fluorescent immunochromatographic method and kit for quantitatively detecting cardiac troponin T in view of the shortcomings of low sensitivity and inaccurate quantification in detecting cardiac troponin T in the prior art

Method used

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  • Fluorescence immunochromatographic assay and kit for quantitatively detecting cardiac troponin T
  • Fluorescence immunochromatographic assay and kit for quantitatively detecting cardiac troponin T
  • Fluorescence immunochromatographic assay and kit for quantitatively detecting cardiac troponin T

Examples

Experimental program
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Effect test

Embodiment 1

[0069] Example 1: Quantitative detection of cTnT by covalently cross-linking antibodies on quantum dots and using direct pre-moisture immunochromatography

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

[0071] Quantum dots with an emission wavelength of 650 nm were mixed with 1 mg / mL cTnT monoclonal antibody and mixed in freshly prepared N-hydroxysuccinimide (NHS, 1 mg / mL) and carbodiimide hydrochloride (EDC, 1 mg / mL) at room temperature for 4-5 hours, adding 1 mol / L glycine to block, and separating and purifying with a chromatographic column or column to obtain cTnT monoclonal antibody-modified quantum dots. Similarly, quantum dots modified with rabbit IgG were obtained. The fluorescence emission wavelength of the quantum dots modified by cTnT antibody is 650 nm, and the fluorescence emission wavelength of the quantum dots modified by rabbit IgG is 570 nm.

[0072] (2) Construction of the kit

[0073] Mix two quantum dot markers in a ratio of 1:1, and add bovin...

Embodiment 2

[0085] Example 2: Quantitative detection of cTnT by using biotin-avidin system to modify antibodies on quantum dots and indirect pre-run immunochromatography

[0086] (1) Modification of quantum dots and antibodies

[0087] First, 1 mL of cTnT monoclonal antibody (1 mg / mL) was fully dialyzed with sodium bicarbonate buffer pH 9.0, and 20-120 μl of N-hydroxysuccinimide biotin ester freshly prepared in dimethyl sulfoxide (DMSO) was added. (NHSB, 1 mg / mL), react at room temperature in the dark for 4 h. Add 1mol / L NH 4 Cl, shake and react at room temperature for 10 min, then put the solution in a dialysis bag, dialysis and purification overnight, and concentrate using an ultrafiltration centrifuge tube to prepare the desired concentration, and the obtained is the biotinylated cTnT monoclonal antibody.

[0088] The streptavidin-modified quantum dots with an emission wavelength of 900 nm and the biotinylated cTnT monoclonal antibody were mixed and reacted in a ratio of 1:3 to 1:12 ...

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Abstract

The invention discloses a fluorescence immunochromatographic assay and kit for quantitatively detecting cardiac troponin T (cTnT). The fluorescence immunochromatographic assay for quantitatively detecting cTnT realizes fluorescence quantitative detection on the basis of optimizing various constituent parts of test paper by using excellent fluorescence characteristic of quantum dots and combining a bicolor marking technology and an immunochromatographic technology. Compared with the conventional colloidal gold immunochromatographic assay, the fluorescence immunochromatographic assay disclosed by the invention has the advantages of good marking stability, low non-specificity, high sensitivity, wide linear range and quantifying accuracy. The fluorescence immunochromatographic kit disclosed by the invention is used for carrying out quantitative detection on the cTnT and detecting whole blood, blood serum and blood plasma samples and is suitable for different levels of hospitals and particularly good for wide popularization in primary hospitals and clinics.

Description

technical field [0001] The invention relates to the field of medical examination, in particular to an early, sensitive and specific quantitative detection of acute myocardial infarction by utilizing cardiac troponin T-related immune response and fluorescence immunochromatography technology. Background technique [0002] Currently, 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 for chest pain every year. The number of deaths has exceeded 500,000. In my country, the prevalence, incidence and risk factors of cardiovascular disease are on the rise. According to statistics in recent years, the mortality rate of cardiovascular disease accounts for 40% of the population death, which is higher than that of European and American countries. It is a country with high incidence of cardiovascular disease. Therefore, on...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68
Inventor 王秀利
Owner SHENZHEN KANGMEI BIOTECH
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