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Joint detection method of cardio-cerebral-vascular disease (CCVD) biomarkers and diagnostic kit

A technology of cardiovascular and cerebrovascular diseases and biomarkers, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of poor repeatability, limited detection throughput, and insufficient sensitivity of solid-phase biochip technology, and achieve high Sensitivity, rapid detection, and good stability

Inactive Publication Date: 2011-06-29
邵棠
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these technologies have limitations in detection throughput, cumbersome operation, poor sensitivity, and cannot really meet the needs of clinical diagnosis and detection.
However, solid-phase biochip technology has the disadvantages of poor reproducibility, insufficient sensitivity and cumbersome operation.

Method used

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  • Joint detection method of cardio-cerebral-vascular disease (CCVD) biomarkers and diagnostic kit
  • Joint detection method of cardio-cerebral-vascular disease (CCVD) biomarkers and diagnostic kit
  • Joint detection method of cardio-cerebral-vascular disease (CCVD) biomarkers and diagnostic kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1: Liquid-phase chip combined parallel detection method for three biomarkers of cardiovascular and cerebrovascular diseases

[0048] The specific detection method includes the following steps:

[0049] Choose 3 kinds of microspheres numbered 15, 37, and 40

[0050] 1. Activation of desired microspheres

[0051] 1.1 Vortex the microsphere storage solution at full speed for at least 3 minutes to form a uniform microsphere suspension;

[0052] 1.2 Weigh 10 mg of EDC and S-NHS into two centrifuge tubes;

[0053] 1.3 Dissolve in deionized water to make the final concentration 50mg / ml;

[0054] 1.4 Take 1ml of the microsphere suspension and centrifuge at 10000g for 3min, carefully remove the supernatant;

[0055] 1.5 Add 80 μl of activation buffer to resuspend the microspheres;

[0056] 1.6 Add 10 μl of EDC solution (50 mg / ml) and 10 μl of S-NHS solution (50 mg / ml) respectively, mix well, and incubate at room temperature (15-25° C.) in the dark and shake for 20 m...

Embodiment 2

[0103] Example 2: Liquid-phase chip combined detection method for 5 biomarkers of cardiovascular and cerebrovascular diseases

[0104] The specific detection method includes the following steps:

[0105] Select 5 kinds of microspheres numbered 15, 21, 33, 37, and 40

[0106] 1-3 steps are the same as in Example 1.

[0107] 4. Configuration of Antigen Standards

[0108] IL-6, TNFα were prepared according to the concentration of 312.5, 62.5, 12.5, 2.5, 0.5, 0.1, 0pg / ml, sICAM1 and sE-Selectin were prepared according to the concentration of 3125, 625, 125, 25, 5, 1, 0ng / ml For preparation, sCD40L was prepared at concentrations of 31.25, 6.25, 1.25, 0.25, 0.05, 0.01, and 0 ng / ml, and the marker mixtures were labeled as STD6, ​​STD5, STD4, STD3, STD2, STD1, and STD0.

[0109] 5. Preparation of microsphere mixture (mixture I) coupled with capture antibody

[0110] Take the microspheres coated with capture antibodies of five cardiovascular and cerebrovascular biomarkers, as follo...

Embodiment 3

[0133] Example 3: Liquid-phase chip combined detection method for 6 biomarkers of cardiovascular and cerebrovascular diseases

[0134] The specific detection method includes the following steps:

[0135] Select 6 kinds of microspheres numbered 11, 15, 21, 33, 37, and 40

[0136] 1-3 steps are the same as in Example 1.

[0137] 4. Configuration of Antigen Standards

[0138] SAA was prepared according to the concentration of 312.5, 62.5, 12.5, 2.5, 0.5, 0.1, 0ug / ml, IL-6 and TNFα were prepared according to the concentration of 312.5, 62.5, 12.5, 2.5, 0.5, 0.1, 0pg / ml, sICAM and sE-Selectin was prepared according to the concentration of 3125, 625, 125, 25, 5, 1, 0ng / ml, sCD40L was prepared according to the concentration of 31.25, 6.25, 1.25, 0.25, 0.05, 0.01, 0ng / ml, and the marker mixture Labeled as STD6, ​​STD5, STD4, STD3, STD2, STD1, STD0.

[0139] 5. Preparation of microsphere mixture (mixture I) coupled with capture antibody

[0140] Take the microspheres coated with c...

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PUM

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Abstract

The invention discloses a joint detection method of cardio-cerebral-vascular disease (CCVD) biomarkers and a diagnostic kit. The joint parallel detection method is characterized in that a plurality of the biomarkers in the same sample can be detected at one time, namely, a tetrad complex of 'detection antibodies for signal marks-the CCVD biomarkers-capture antibodies-beads' is formed by preparing a liquid chip, and then existence and content of various different CCVD biomarkers in the sample to be detected are determined by detecting mark signals of different beads in the tetrad complex. The invention further discloses composition and application of the diagnostic kit. The method and the kit have the outstanding advantages of high throughput, high sensitivity, high specificity, rapid and accurate detection and the like, and can be used for qualitatively and quantitatively detecting the plurality of the CCVD biomarkers at the same time.

Description

technical field [0001] The invention relates to an in vitro diagnostic detection method and a diagnostic kit thereof, in particular to a liquid-phase chip combined parallel detection method of biomarkers of cardiovascular and cerebrovascular diseases and a diagnostic kit thereof. Background technique [0002] Cardiovascular and cerebrovascular diseases are one of the major diseases and leading causes of death in China's urban population and western developed countries, mainly including cardiovascular diseases represented by coronary heart disease and hypertension, as well as ischemic cerebrovascular diseases and hemorrhagic cerebrovascular diseases sick. Ischemic cerebrovascular disease, also known as acute ischemic cerebrovascular syndrome (AICS), includes cerebral infarction (CI) and transient ischemic attack (TIA); hemorrhagic cerebrovascular disease includes cerebral hemorrhage and arachnoid Inferior hemorrhage. The joint detection of biomarkers of cardiovascular and c...

Claims

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Application Information

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IPC IPC(8): G01N33/546
Inventor 邵棠黄梅
Owner 邵棠
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