Chemiluminescent detection microfluidic chip and chemiluminescent detection microfluidic chip system and application thereof
A chemiluminescence detection, microfluidic chip technology, applied in the application of chemiluminescence detection, chemiluminescence detection microfluidic chip system, chemiluminescence detection microfluidic chip field, can solve single sample detection accuracy decline, capillary Problems such as uneven force and large changes in chip structure have achieved the effect of strong detection repeatability, wide linear range, and avoiding interference and pollution problems
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preparation example Construction
[0083] The preparation method of the chemiluminescent detection microfluidic chip provided by the present invention preferably includes the following steps:
[0084] Step 1) preload the reagents required for the chemiluminescence reaction into the reagent pouch 21, and seal the sealing port 212 of the reagent pouch 21 by means of thermoplastic sealing; and place it on the chip through the positioning hole 211 At the specific liquid storage structure of the middle layer structure of the system, the reagent pouches 21 for each liquid storage are further fixed on the middle layer structure of the chip system by means of thermal plastic sealing.
[0085] Step 2) Bonding the lower structure to the lower part of the upper middle structure, the bonding methods include but not limited to ultrasonic melting, adhesive, ultraviolet curing, thermoplastic sealing and other methods.
[0086] Step 3) Attaching the upper structure to the upper middle structure, the attachment methods include ...
Embodiment 1
[0097] This example is used to illustrate the use of magnetic particle chemiluminescence to detect C-reactive protein (CRP) in whole blood
[0098] The enzyme-labeled antibody used in this example is horseradish peroxidase-labeled CRP (HRP-CRP), the size of the magnetic particles in the magnetic particle-labeled CRP is about 5 μm, and the cleaning solution contains 0.2 volume % BSA, 0.5 volume % tween -20 phosphate buffer solution (pH 7.5), the first luminescent base solution is an acidic solution containing luminol, and the second luminescent base solution is an alkaline solution containing benzene derivatives.
[0099] (1) Assembly of chip system
[0100] The above-mentioned reaction reagents are prepacked in the reagent pouch 21, and the sealing port 212 of the reagent pouch 21 is sealed by means of thermoplastic sealing (heating time 30s); and placed in the middle layer structure of the chip system through the positioning hole 211 At the specific liquid storage structure,...
Embodiment 2
[0108] This example is used to illustrate the detection of C-reactive protein (CRP) in whole blood by the coated antibody chemiluminescence method
[0109] The enzyme-labeled antibody used in this embodiment is horseradish peroxidase-labeled CRP (HRP-CRP), and the cleaning solution is a phosphate buffer (pH 7.5) containing 0.2 volume % BSA and 0.5 volume % tween-20. The first luminescent base liquid is an acidic solution containing luminol, and the second luminescent base liquid is an alkaline solution containing benzene derivatives. The coated antibody is a C-reactive protein solution, which is injected into the reaction unit 3 by spotting the sample, and baked at 40°C for 3 hours after spotting, so that the CRP protein spotted at the reaction unit 3 is adsorbed and fixed. in the reaction chamber.
[0110] The detection process is the same as in Example 1.
[0111] The results showed that the minimum detection limit of CRP concentration detection by the coating antibody met...
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