Magnetic particle micro-fluidic biochip for cardio-pulmonary function marker and detection method

A biochip and cardiopulmonary function technology, which is applied in biological testing, biomaterial analysis, chemical instruments and methods, etc., can solve the problems of inability to miniaturize equipment and POCT, reduce detection precision, detection sensitivity and specific interference, etc. , to avoid missed detection and false negative phenomenon, increase rescue time and shorten diagnosis time

Inactive Publication Date: 2018-11-16
微粒云科技(北京)有限公司
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  • Summary
  • 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

Method used

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  • Magnetic particle micro-fluidic biochip for cardio-pulmonary function marker and detection method
  • Magnetic particle micro-fluidic biochip for cardio-pulmonary function marker and detection method
  • Magnetic particle micro-fluidic biochip for cardio-pulmonary function marker and detection method

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Embodiment 1

[0094] The present invention is a cardiopulmonary function marker magnetic particle microfluidic biochip, the biosensor is pre-coated with six cardiopulmonary function markers (CTI / CK-MB / MYO / D-Dime / NT-proBNP / hs-CRP) spots sample monoclonal antibody, and then stick the first layer 30, the second layer 40, the microfluidic channel sealing layer 50 and the whole blood filter layer 60 on the microfluidic channel 11 of the PCB board 10 in sequence, thereby sealing the microfluidic channel 11 , so that the reaction is carried out under airtight 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

[0097] The preparation method of the biochip for detection of six cardiopulmonary function markers (CTI / CK-MB / MYO / D-Dime / NT-proBNP / hs-CRP) of the present invention:

[0098] 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 30nm and 50nm black colloidal superparamagnetic supersensitive magnetic particle mixed suspension, and the antibody / avidin with amino group is coupled. Quantitative detection reagent preparation steps are as follows,

[0099] 1. Preparation of 30nm+50nm superparamagnetic ultrasensitive magnetic particles:

[0100] (1) Preparation of 30nm magnetic beads: Take 0.75ml~2.25ml 4% Fe 3 o 4 Add the solution into 300ml of ultrapure water to make the final concentration of ferric oxide solution 0.0...

Embodiment 3

[0178] The specific steps of the detection of the six markers of cardiopulmonary function of the present invention:

[0179] (1) Take the enterprise quality control product and biochip quantitative detection kit out of the refrigerator, and equilibrate to room temperature;

[0180] (2) Turn on the chip detector, perform system self-test and preheat for 5 minutes;

[0181] (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.

[0182] (4) Adding samples: Use a 100ul micropipette to pipette 25ul of the internal control substances of the six markers of cardiopulmonary function into the corresponding chip loading area, and insert the chip into the detector;

[0183] (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 1...

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Abstract

The invention relates to a magnetic particle micro-fluidic biochip for cardio-pulmonary function markers, and belongs to the technical field of POCT detection. The micro-fluidic biochip comprises a PCB, the PCB is provided with a micro-fluidic channel , a biosensor is arranged in the micro-fluidic channel and comprises a wafer platform and sample application antibodies coated on the wafer platform, the sample application antibodies, magnetic bead coupling antibodies and marker protein can form an immune complex, the concentration of the marker protein is judged by measuring the strength of themagnetic resistance signal on the complex on the wafer platform, and the sample application antibodies are antibodies for the six cardio-pulmonary function markers of CTI, CK-MB, MYO, D-Dime, NT-proBNP and hs-CRP. The micro-fluidic biochip is characterized in tha, magnetic beads have relatively larger specific surface area, more protein/antibodies are combined, and therefore the detection sensitivity is improved. The invention further provides a detection method applying the biochip. The detection method has the advantages that the cardiac markers of myocardial infarction, congestive heart failure and pulmonary embolism can be simultaneously and quantitatively detected, in addition, the lower limit of detection is improved, and the missing detection and false negative phenomena are effectively avoided.

Description

technical field [0001] The invention relates to a biochip, which is mainly used for detecting six markers of heart and lung function 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, ...

Claims

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Application Information

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IPC IPC(8): G01N33/68B01L3/00
CPCB01L3/5027B01L2200/10G01N33/6893G01N2800/32
Inventor 张永勇
Owner 微粒云科技(北京)有限公司
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