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Biomarker diagnostic kit for acute coronary syndrome

A technology of biomarkers and coronary arteries, which is applied in biological testing, chemiluminescence/bioluminescence, and analysis by making materials undergo chemical reactions, can solve the problem of poor repeatability, insufficient sensitivity, and poor sensitivity of solid-phase biochip technology. And other problems, to achieve high sensitivity, good stability, rapid detection effect

Active Publication Date: 2013-09-18
MEDI GENETECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these technologies can only detect one marker at a time, and the operation is cumbersome and the sensitivity is poor, which 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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  • Biomarker diagnostic kit for acute coronary syndrome
  • Biomarker diagnostic kit for acute coronary syndrome
  • Biomarker diagnostic kit for acute coronary syndrome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1: Liquid-phase chip combined parallel detection method for 3 kinds of acute coronary syndrome markers

[0045] The specific detection method includes the following steps:

[0046] 1. Activation of required microspheres:

[0047] 1.1 Vortex the microsphere storage solution at full speed for at least 3 minutes to form a uniform microsphere suspension;

[0048] 1.2 Weigh 10mg of EDC and S-NHS into two centrifuge tubes;

[0049] 1.3 Dissolve with deionized water to make the final concentration 50mg / ml;

[0050] 1.4 Take 1ml of microsphere suspension at 10000g and centrifuge for 3min, carefully remove the supernatant;

[0051] 1.5 Add 80μl of activation buffer to resuspend the microspheres;

[0052] 1.6 Add 10μl of EDC solution (50mg / ml) and 10μl of S-NHS solution (50mg / ml) respectively, mix well, at room temperature (15-25℃), avoid light, and incubate with shaking for 20min.

[0053] 2. The corresponding capture antibody is coupled to the activated microsphere

[0054] 2.1 Dilute...

Embodiment 2

[0097] Example 2: Liquid-phase chip combined parallel detection method for 6 kinds of acute coronary syndrome markers The specific detection method, steps 1-3 are the same as in Example 1.

[0098] 4. Configuration of antigen standard

[0099] cTnT, cTnI are prepared at the concentration of 31.25, 6.25, 1.25, 0.25, 0.05, 0.01, 0ng / ml, and CK-MB and H-FABP are prepared at the concentration of 312.5, 62.5, 12.5, 2.5, 0.5, 0.1, 0ng / ml Preparation, Myo and MPO were prepared at the concentration of 3125, 625, 125, 25, 5, 1, 0 ng / ml, and the marker mixture was labeled STD6, ​​STD5, STD4, STD3, STD2, STD1, STD0.

[0100] 5. Preparation of microsphere mixture (I mixture) coupled with capture antibody

[0101] Take the capture antibody microspheres coated with 6 ACS markers, as follows: cTnT capture antibody microsphere 11, cTnI capture antibody microsphere 15, CK-MB capture antibody microsphere 21, H-FABP capture antibody microsphere 33. Myo capture antibody microsphere 35, MPO capture antib...

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Abstract

The invention discloses a biomarker detection method and a diagnostic kit for acute coronary syndrome. The detection method is characterized in that: a plurality of biomarkers in the same sample can be detected once, namely, a biotin-marked detection antibody-acute coronary syndrome biomarker-capture antibody-bead tetragenous complex is formed by the preparation of a liquid chip, and is combined with streptavidin-phycoerythrin (PE) to detect fluorescence signals of different beads, thereby determining the existence of different acute coronary syndrome biomarkers in the sample to be detected and the acute coronary syndrome biomarker content of the sample. The invention also discloses components of the diagnostic kit. The method and the kit provided by the invention have the advantages of high sensitivity, high flux, high detection speed, detection accuracy and the like, and can simultaneously realize the qualitative and quantitative detection of the plurality of acute coronary syndrome biomarkers.

Description

technical field [0001] The invention relates to an in vitro diagnostic detection method and a diagnostic kit, in particular to a liquid-phase chip combined parallel detection method and a diagnostic kit for biomarkers of various acute coronary syndromes. Background technique [0002] Acute Coronary Syndrome (ACS) is a group of clinical syndromes characterized by instability of coronary atherosclerotic plaques, acute myocardial ischemia as a common feature, and chest pain as a typical symptom. ACS is the main reason for most patients with cardiovascular disease to see a doctor and die suddenly, which seriously endangers human life and health. ACS biomarkers (hereinafter referred to as "ACS markers") can not only accurately diagnose acute coronary syndrome and evaluate its severity, but also guide clinical treatment and efficacy monitoring, and judge the prognosis of acute coronary syndrome. Therefore, the combined parallel detection of various types of ACS markers has become...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/76G01N21/64G01N33/68G01N33/70
Inventor 邵棠孙黎吴杰
Owner MEDI GENETECH
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