Microfluidic chip based on micro liquid drops and detection system
A microfluidic chip and microdroplet technology, applied in the field of clinical in vitro detection and biotechnology, can solve the problems of low repeatability, high cost, and large sample demand.
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Embodiment 1
[0221] Detection of BNP in human blood samples by chemiluminescent method
[0222] In this example, using figure 1 The microfluidic chip shown uses chemiluminescent method to detect BNP in human blood samples.
[0223] 1. Experimental method
[0224] 1. Carry out the pretreatment of blood samples and reagents, that is, take human blood samples A and B, and add them into EDTA-anticoagulant blood collection tubes to prevent coagulation.
[0225] 2. The treatment of BNP reagent: the BNP antibody is coated with magnetic beads, and the BNP antibody is labeled with acridine.
[0226] 3. Preparation of chemiluminescent substrate: equal volume of 0.1M HNO 3 , 0.1%H 2 o 2 Luminescence excitation solution A is mixed with 0.25M NaOH, 2% Tween-20 luminescence excitation solution B to form a chemiluminescence excitation substance.
[0227] 4. Sample injection: in figure 1 Add sample A to the dispersed phase inlet 6 shown, in figure 1 Continuous phase mineral oil is added to the con...
Embodiment 2
[0244] Detection of B-type natriuretic peptide (BNP) by double-antibody sandwich method
[0245] In this example, using figure 1 The microfluidic chip shown uses the double-antibody sandwich method to detect BNP in human blood samples.
[0246] 1. Experimental method
[0247] 1. same as embodiment 1, first carry out the pretreatment of blood sample and reagent: get human blood sample A in EDTA anticoagulant tube, to prevent coagulation.
[0248] 2. Treat the two antibodies of the double-antibody sandwich method: fluorescently labeled antibody 1 and magnetic bead-coupled antibody 2.
[0249] 3. Sample injection: in figure 1 Add sample A to the dispersed phase inlet 6 shown, in figure 1 Add the continuous phase mineral oil in the continuous phase inlet 1 shown, at this moment the control valve 5 is closed, and the control valve 7 is opened. figure 1 The shown sample outlet 28 and waste liquid port 4 give negative pressure as a power source, and the design of the sample inle...
Embodiment 3
[0264] Construction of standard curve for detection of B-type natriuretic peptide (BNP) by double-antibody sandwich method
[0265] In this example, for the detection of B-type natriuretic peptide (BNP) by the double-antibody sandwich method based on the immune response, the results in different extraction processes were compared, and a standard curve was prepared for it. In addition, based on the test results, the cleaning effects were compared.
[0266] The method is as follows: first prepare a series of concentrations of BNP calibrator; using the same calibrator, the number of micro-droplets cleaned during (a) extraction is twice that of the sample droplets; and (b) the number of micro-droplets cleaned during extraction is In the case of twice the sample droplet, the determination is carried out separately, the results of different extraction processes are compared, and a standard curve is made.
[0267] The comparison results are as Image 6 , Figure 7 , and shown in T...
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