Magnetic particle microfluidic biochip for four inflammation markers and detection method
A biochip and marker technology, applied in biological testing, chemical instruments and methods, material testing products, etc., can solve the problem of inability to miniaturize and POCT equipment, detection sensitivity and specificity interference, and inability to achieve bedside detection, etc. problems, to avoid missed detection and false negatives, improve sensitivity, and increase rescue time.
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Embodiment 1
[0101]The invention relates to a magnetic particle microfluidic biochip for four markers of inflammation. The biosensor is pre-coated with monoclonal antibodies for the four markers of inflammation (PCT / IL-6 / CRP / SAA), and then the first The 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 under airtight conditions , to achieve the effect of 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 closed pipeline, At ...
Embodiment 2
[0104] The preparation method of the biochip for detection of four markers of inflammation (PCT / IL-6 / CRP / SAA) of the present invention:
[0105] The method of coupling superparamagnetic nano-magnetic particles is adopted. The superparamagnetic nano-magnetic particles include mixed magnetic beads with a diameter of 30nm and a diameter of 50nm. The coupling mechanism of the nano-magnetic particles is through covalent bonding. The effect of different concentrations 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,
[0106] 1. Preparation of 30nm+50nm superparamagnetic ultrasensitive magnetic particles:
[0107] (1) Preparation of 30nm magnetic beads: Take 0.75ml~2.25ml 4% Fe 3 o 4 Add the solution to 300ml of ultra-pure water to make the final concentration of ferric oxide solution...
Embodiment 3
[0185] The specific steps of the detection of the four markers of inflammation of the present invention:
[0186] (1) Take the enterprise quality control product and biochip quantitative detection kit out of the refrigerator, and equilibrate to room temperature;
[0187] (2) Turn on the chip detector, perform system self-test and preheat for 5 minutes;
[0188] (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.
[0189] (4) Adding samples: use a 100ul micropipette to respectively pipette 25ul of the internal control products of the four markers of inflammation into the corresponding chip loading area, and insert the chip into the detector;
[0190] (5) Detection: Select the corresponding inflammatory marker detection module of the detector to start the detection, and the device will automatically read the results in 15 to 20 minutes...
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