Biochip and detection method for detecting five-component markers of myocardium

A biochip and marker technology, applied in biological testing, measurement devices, material inspection products, etc., can solve the problems of inability to miniaturize and POCT equipment, reduce detection precision, and have a large impact on characteristic sensitivity, so as to avoid leakage. The effect of detecting and false negative phenomenon, shortening diagnosis time, and broadening the linear range

Inactive Publication Date: 2018-11-20
微粒云科技(北京)有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The first-generation detection technology competition method (radioimmunoassay, RIA): There are many factors affected, and the RIA sample requires a large amount. Because extraction is usually required before detection, it not only increases the operation steps, but also because the recovery rate of extraction is usually Only 80% to 90%, the coefficient of variation (CV) is relatively large, which reduces the precision of detection;
[0009] The second-generation detection method non-competitive method (radioimmunoassay, RIA): immunoassay using double-antibody sandwich, no extraction, high antibody specificity, its sensitivity, precision and specificity are better than competitive methods, but time-consuming Still long and difficult to apply to automation;
[0010] The third-generation detection method enzyme immunoassay, stationary phase immunochromatography: the double antibody plus core method immunoassay solid phase chroma

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  • Biochip and detection method for detecting five-component markers of myocardium
  • Biochip and detection method for detecting five-component markers of myocardium
  • Biochip and detection method for detecting five-component markers of myocardium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0093] The present invention is a biochip for detection of five myocardial markers, the biosensor is pre-coated with five myocardial markers (CTI / CK-MB / MYO / D-Dime / NT-proBNP) spotted monoclonal antibody, Then the first layer 30, the second layer 40, the microfluidic channel sealing layer 50, and the whole blood filter layer 60 are pasted on the microfluidic channel 11 of the PCB board 10 in sequence, thereby sealing the microfluidic channel 11, so that the reaction can be carried out in a sealed environment. It is carried out under certain conditions to achieve the effect of a micro-full laboratory. The first layer 30 is a biosensor airtight layer, containing the sampling hole 31 and the waste liquid hole 32; The pump guides the sample to be tested to flow in one direction from the mixing area 41 to the detection area (where the biosensor is located) and the waste liquid area 44; the microfluidic channel sealing layer 50 is the third layer to ensure that the sample flows in a c...

Embodiment 2

[0096] The preparation method of the biochip for the detection of five myocardial markers (CTI / CK-MB / MYO / D-Dime / NT-proBNP) of the present invention:

[0097] Using the coupling method of superparamagnetic nano-magnetic particles, the superparamagnetic nano-magnetic particles include mixed magnetic beads with a diameter of 30nm and a diameter of 50nm. The reaction mechanism of the nano-magnetic particles is covalent bonding. The concentration of sodium acetate is reduced to a mixed suspension of 30nm and 50nm black colloidal superparamagnetic supersensitive magnetic particles, and the antibody / avidin with amino groups is coupled. Quantitative detection reagent preparation steps are as follows,

[0098] 1. Preparation of 30nm+50nm superparamagnetic ultrasensitive magnetic particles:

[0099] (1) Preparation of 30nm magnetic beads: Take 0.75ml~2.25ml 4% Fe 3 o 4 Add the solution to 300ml of ultrapure water to make the final concentration of the iron oxide solution 0.01% to 0.0...

Embodiment 3

[0175] The specific steps of detection of the five myocardial markers (CTI / CK-MB / MYO / D-Dime / NT-proBNP) of the present invention:

[0176] (1) Take the enterprise quality control product and biochip quantitative detection kit out of the refrigerator, and equilibrate to room temperature;

[0177] (2) Turn on the chip detector, perform system self-test and preheat for 5 minutes;

[0178] (3) Tear open the required aluminum foil bag containing the chip and take out the biochip, and mark the type and concentration of the quality control substance tested in the ID information position.

[0179] (4) Adding samples: Use a 100ul micropipette to pipette 25ul of the five internal control substances of the myocardium into the corresponding chip injection holes, and insert the chip into the detector;

[0180] (5) Detection: Select the corresponding myocardial marker detection module of the detector to start the detection, and the device will automatically read the results within 15 to 20 ...

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Abstract

The invention relates to a biochip for detecting five-component markers of myocardium, and belongs to the technical field of POCT (Point-of-care Testing) detection. The biochip comprises a biosensor;the biosensor comprises a wafer platform and a point-like antibody coated on the wafer platform; the point-like antibody can form an immune complex with a magnetic bead-conjugated antibody and a marker protein, and the marker protein concentration can be determined by measuring the magneto resistance signal intensity on the complex on the biosensor. In the technical scheme, magnetic beads have a relatively large specific surface area, and combined with more antibodies, the detection sensitivity is improved, and the specificity is higher than that of a conventional chemiluminescence method. Theinvention further provides a detection method applicable to the biochip; by adopting the detection method, the detection range of myocardial markers is improved, cardiac markers of myocardial infarction, heart failure and pulmonary embolism can be quantitatively detected, the linear range of detection is expanded, and missing detection and false-negative phenomenon can be effectively avoided; andthe detection method is short in detection time, safe, pollution-free, and suitable for ambulance, emergency department, ICU (intensive care unit)and bedside detection.

Description

technical field [0001] The invention relates to a biochip, which is mainly used for detecting five myocardial markers in blood samples; the invention also provides a detection method using the biochip, which belongs to the technical field of POCT detection. Background technique [0002] Cardiovascular disease is a serious disease that endangers human health and life, and has become the number one killer of human health in the 21st century. According to the "China Cardiovascular Disease Report 2015" and "China Cardiovascular Disease Report 2016" compiled by the National Center for Cardiovascular Diseases, the number of patients with cardiovascular diseases in my country is still growing rapidly. According to statistics, the number of patients with cardiovascular disease was 290 million in 2014, and this number has climbed to 290 million by 2015. According to the report estimates, about 3.5 million people die of cardiovascular disease in my country every year, accounting for ...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/6854
Inventor 张永勇
Owner 微粒云科技(北京)有限公司
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