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A liquid chip kit for acute coronary syndrome and its preparation method

A technology for coronary syndrome and liquid phase chip, which is applied to the field of liquid chip kit and its preparation, can solve the problems of complex preparation method of the kit, poor application effect, poor stability and the like, and achieves simple and easy preparation method, Small sample size and good stability

Inactive Publication Date: 2015-11-18
吴宗贵 +2
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0006] The purpose of the present invention is to provide an improved liquid-phase chip kit for acute coronary syndrome and its preparation method, which can overcome the diagnostic kits in the prior art and focus on improving the detection of ACS by means of peripheral serum markers. In terms of diagnostic sensitivity and specificity, there is still a gap in the relevant kits for improving the risk stratification and prognosis of patients. At the same time, the existing kits have some shortcomings such as complex preparation methods, poor stability, and poor application effects.

Method used

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  • A liquid chip kit for acute coronary syndrome and its preparation method
  • A liquid chip kit for acute coronary syndrome and its preparation method
  • A liquid chip kit for acute coronary syndrome and its preparation method

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preparation example Construction

[0039] A method for preparing a liquid chip kit for acute coronary syndrome, characterized in that: the preparation method includes the following steps:

[0040] 1) Prepare the corresponding antibody-coated microspheres according to the composition of the above kit:

[0041] Select different microspheres respectively, and suspend them by vortexing or ultrasonic for 20-25 seconds;

[0042] After 50 μl of microspheres are activated, the centrifugation condition is ≥8000g, and centrifuged for 1-2 minutes;

[0043] Aspirate the supernatant, resuspend the microspheres in 250 μl of 50 mM MES solution, vortex for 25-30 seconds, sonicate for 25-30 seconds, and collect the activated microspheres by centrifugation, the centrifugation condition is ≥8000g, centrifuge for 1-2 minutes ;Repeat this step once;

[0044] Aspirate the supernatant, resuspend the microspheres in 100 μl of 50 mM MES solution, vortex for 25-30 seconds, sonicate for 25-30 seconds, add 1 μg of antibody, and make up th...

Embodiment 1

[0070] Preparation of liquid chip and antigen detection for risk stratification and prognosis judgment in patients with acute coronary syndrome

[0071] The capture antibody of the present invention is a monoclonal antibody that can specifically bind to ox-LDL, sCD40L, MMP-9, TF, omentin and esRAGE respectively; Monoclonal or polyclonal antibodies specifically binding to MMP-9, TF, omentin and esRAGE.

[0072] The capture antibodies and detection antibodies of "ox-LDL, sCD40L, MMP-9, TF, omentin, and esRAGE" used in this example were purchased from Abcam, Santacruz, Invitrogen, and Millipore, respectively.

[0073] Microspheres with different numbers (surface carboxyl modification) and streptavidin-phycoerythrin were purchased from Luminex, USA.

[0074] EDC, S-NHS and S-NHS-Biotin were purchased from Pierce Company.

[0075] In the present embodiment, the formula of described various solutions is as follows:

[0076] 1. 50mM MES buffer (pH5.0) formula (500ml): Dissolve 4.8...

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Abstract

The invention relates to a liquid-phase chip kit for acute coronary syndrome and a preparation method for the same. A liquid-phase chip mainly comprises microspheres coated with oxidized low density lipoprotein (ox-LDL), soluble CD40 ligand (sCD40L), matrix metallopeptidase 9 (MMP-9), tissue factor (TF), omentin and endogenous secretory receptor of advanced glycation end products (esRAGE) capture antibody respectively, detection antibodies labelled by biotin, and streptavidin phycoerythrin. The liquid-phase chip provided by the invention has the advantages of being high in detection efficiency, low in the quantity of the needed samples, strong in specificity, high in sensitivity, rapid and accurate in detection, and the like. Simultaneously, the chip reflects a protective factor level and an injury factor level in an organism, so that the risk stratification and prognosis conditions of a patient can be much comprehensively assessed. Simultaneously, the preparation method disclosed by the invention is simple and practicable, and good in stability, various process parameters such as the quantities of the microspheres and the antibodies, and the reaction process in the technical scheme of the preparation method are obtained on the basis of lots of experiments, and are the optimal parameter values of the preparation process.

Description

Technical field: [0001] The invention relates to the technical field of in vitro kits, in particular to a liquid phase chip kit for acute coronary syndrome and its preparation method. Background technique: [0002] Atherosclerosis (AS) vascular disease has become the "number one killer" that threatens the health of Chinese people. With the development of my country's economy, the incidence rate is increasing year by year. The annual medical expenses for such diseases in my country are as high as 100 billion Strengthening the prevention and treatment of AS vascular diseases and their complications has become the main task of the prevention and treatment of major diseases in our country. [0003] Acute coronary syndrome (ACS) caused by AS vascular disease is the most common and serious clinical manifestation, and the rupture of vulnerable coronary plaque followed by thrombosis is the main pathological basis of its occurrence. At present, coronary angiography, which is the "gol...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577
Inventor 吴宗贵梁春贺治青张家友潘晓明樊民李玫丁茹甄奕伍锋朱琳王新刘焕波
Owner 吴宗贵
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