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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, increasing the sensitivity, and improving the detection sensitivity

Active Publication Date: 2012-07-11
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

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Embodiment 1

[0069] Example 1: Quantitative detection of cTnT by modifying antibodies to quantum dots by covalent cross-linking and using direct pre-wetting immunochromatography

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

[0071] Quantum dots with an emission wavelength of 650nm were mixed with 1mg / mL cTnT 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 using a chromatographic column or chromatographic column to separate and purify to obtain cTnT monoclonal antibody modified quantum dots. Similarly, rabbit IgG-modified quantum dots were obtained. The fluorescence emission wavelength of the cTnT antibody-modified quantum dot is 650nm, and the fluorescence emission wavelength of the rabbit IgG-modified quantum dot is 570nm.

[0072] (2) Construction of the kit

[0073] Mix the two kinds of quantum dot...

Embodiment 2

[0085] Example 2: Quantitative detection of cTnT by modifying antibody to quantum dots with biotin-avidin system and using indirect pre-wetting immunochromatography

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

[0087] First, 1 mL of cTnT 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 cTnT monoclonal antibody.

[0088] Streptavidin-modified quantum dots with an emission wavelength of 900nm and biotinylated cTnT monoclonal antibody were mixed and reacted at a ratio of 1:3-1:12 for 30-60 mi...

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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 testing, 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] At present, cardiovascular disease has become the biggest potential killer of adults in the world. 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. The number of deaths 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...

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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