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Joint detection method for heart cerebrovascular disease-related protein marker and diagnostic kit thereof

A technology of cardiovascular and cerebrovascular diseases and protein markers, which is applied in the field of liquid phase chip combined with parallel detection method and its diagnostic kit, can solve the problem of poor reproducibility of solid phase biochip technology, limitation of detection throughput, insufficient sensitivity, etc. problems, to achieve high sensitivity, rapid detection, and good stability

Inactive Publication Date: 2012-02-15
MEDI GENETECH
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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 for heart cerebrovascular disease-related protein marker and diagnostic kit thereof
  • Joint detection method for heart cerebrovascular disease-related protein marker and diagnostic kit thereof
  • Joint detection method for heart cerebrovascular disease-related protein marker and diagnostic kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1: Liquid-phase chip joint detection method for two kinds of protein markers related to cardiovascular and cerebrovascular diseases

[0049] The specific detection method includes the following steps:

[0050] Select 2 kinds of microspheres numbered 11 and 15

[0051] 1. Activation of desired microspheres

[0052] 1.1 Vortex the microsphere storage solution at full speed for at least 3 minutes to form a uniform microsphere suspension;

[0053] 1.2 Weigh 10 mg of EDC and S-NHS into two centrifuge tubes;

[0054] 1.3 Dissolve in deionized water to make the final concentration 50mg / ml;

[0055] 1.4 Take 1ml of the microsphere suspension and centrifuge at 10000g for 3min, carefully remove the supernatant;

[0056] 1.5 Add 80 μl of activation buffer to resuspend the microspheres;

[0057] 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 2...

Embodiment 2

[0110] Example 2: Liquid-phase chip combined detection method for three kinds of protein markers related to cardiovascular and cerebrovascular diseases

[0111] The specific detection method includes the following steps:

[0112] Select 3 kinds of microspheres numbered 21, 33, and 37

[0113] 1-3 steps are the same as in Example 1.

[0114] 4. Configuration of Antigen Standards

[0115] D-D was prepared according to the concentration of 31250, 6250, 1250, 250, 50, 10, 0ng / ml, PAI-1 and tPA were prepared according to the concentration of 312.5, 62.5, 12.5, 2.5, 0.5, 0.1, 0ng / ml, and the markers The mixtures are labeled as STD6, ​​STD5, STD4, STD3, STD2, STD1, STD0, respectively.

[0116] 5. Preparation of the microsphere mixture (I mixture) coupled with the capture antibody.

[0117] Take the microspheres coated with the capture antibodies of three kinds of cardiovascular and cerebrovascular-related protein markers, as follows: D-D capture antibody microspheres 21, PAI-1 ca...

Embodiment 3

[0144] Example 3: Liquid chip combined detection method for 5 kinds of protein markers related to cardiovascular and cerebrovascular diseases

[0145] The specific detection method includes the following steps:

[0146] Select 5 kinds of microspheres numbered 11, 15, 21, 33, and 37

[0147] 1-3 steps are the same as in Example 1.

[0148] 4. Configuration of Antigen Standards

[0149] D-D is prepared according to the concentration of 31250, 6250, 1250, 250, 50, 10, 0ng / ml, PAI-1, LEP and tPA are prepared according to the concentration of 312.50, 62.50, 12.50, 2.50, 0.50, 0.10, 0ng / ml, ApoB was prepared at concentrations of 31.25, 6.25, 1.25, 0.25, 0.05, 0.01, and 0 mg / ml, and the marker mixtures were labeled as STD6, ​​STD5, STD4, STD3, STD2, STD1, and STD0, respectively.

[0150] 5. Preparation of microsphere mixture (mixture I) coupled with capture antibody

[0151] Take the microspheres coated with capture antibodies of five kinds of protein markers related to cardiovas...

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Abstract

The invention discloses a joint detection method for heart cerebrovascular disease-related protein markers and a diagnostic kit thereof. A joint parallel detection method is characterized in that: a plurality of protein markers of the same sample can be detected at one time, i.e., a quadruple complex consisting of a signal marking detection antibody, a heart cerebrovascular disease-related protein marker, a capture antibody and microspheres is formed by preparing a liquid-phase chip, and the presence and content of various different heart cerebrovascular disease-related protein markers in a sample to be detected are determined by detecting marking signals of different microspheres in the quadruple complex. The invention further discloses constituents of the diagnostic kit and application of the diagnostic kit. The method and the kit provided by the invention have the outstanding advantages of high flux, high sensitivity, high specificity, quick and accurate detection and the like, and can be used for qualitatively and quantitatively detecting a plurality of heart cerebrovascular disease-related protein markers simultaneously.

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 for protein markers related to 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 (also known as cerebral infarction, CI) and transient ischemic attack (TIA); hemorrhagic cerebrovascular disease includes Cerebral hemorrhage and subarachnoid hemorrhage. The combined d...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/532G01N33/533
Inventor 邵棠袁福美陈庆
Owner MEDI GENETECH
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