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Fluorescence immunochromatographic assay and kit for quantitative detection of human cardiac troponin I (cTnI)

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

Active Publication Date: 2014-08-13
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 by the present invention is to provide a fluorescent immunochromatographic method and kit for quantitatively detecting cardiac troponin I in view of the shortcomings of low sensitivity and inaccurate quantification in detecting cardiac troponin I in the prior art

Method used

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  • Fluorescence immunochromatographic assay and kit for quantitative detection of human cardiac troponin I (cTnI)
  • Fluorescence immunochromatographic assay and kit for quantitative detection of human cardiac troponin I (cTnI)
  • Fluorescence immunochromatographic assay and kit for quantitative detection of human cardiac troponin I (cTnI)

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Embodiment 1

[0070] Example 1: Quantitative detection of cTnI by modifying antibodies to quantum dots by covalent cross-linking and using direct pre-wetting immunochromatography

[0071] (1) Modification of quantum dots and antibodies

[0072] Quantum dots with an emission wavelength of 650 nm were mixed with cTnI monoclonal antibody at 1 mg / mL, and mixed with freshly prepared N-hydroxysuccinimide (NHS, 1 mg / mL) and carbodiimide hydrochloride (EDC, 1 mg / mL) at room temperature for 4-5 hours, add 1 mol / L glycine to block, and separate and purify with chromatographic column or chromatographic column to obtain cTnI monoclonal antibody-modified quantum dots. Similarly, rabbit IgG-modified quantum dots were obtained. The fluorescence emission wavelength of the cTnI antibody-modified quantum dot is 650nm, and the fluorescence emission wavelength of the rabbit IgG-modified quantum dot is 570nm.

[0073] (2) Construction of the kit

[0074] Mix the two kinds of quantum dot markers in a ratio o...

Embodiment 2

[0086] Example 2: Quantitative detection of cTnI by modifying antibody to quantum dots with biotin-avidin system and using indirect pre-wetting immunochromatography

[0087] (1) Modification of quantum dots and antibodies

[0088] First, 1 mL of cTnI 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 minutes, then put the solution in a dialysis bag, dialysis and purification overnight, and concentrated with an ultrafiltration centrifuge tube to prepare the required concentration, and the obtained biotinylated cTnI monoclonal antibody.

[0089] Streptavidin-modified quantum dots with an emission wavelength of 900nm and biotinylated cTnI monoclonal antibody were mixed and reacted at a ratio of 1:3-1:12 for 30-60...

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Abstract

The invention discloses a fluorescence immunochromatographic assay and kit for quantitative detection of cardiac troponin I (cTnI). The fluorescence immunochromatographic assay for quantitative detection of the cTnI 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 cTnI, can be used for detecting whole blood, blood serum and plasma samples simultaneously, and is applied to different levels of hospitals and is particularly favored to be widely popularized to primary hospitals and clinics.

Description

technical field [0001] The invention relates to the field of medical examination, in particular to a quantitative detection method for diagnosing acute myocardial infarction by using cardiac troponin I-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. Therefore...

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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