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Chorionic gonadotropin beta subunit microfluidic discs and methods of use thereof
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A chorionic gonadotropin and subunit technology, applied in chemical instruments and methods, laboratory containers, instruments, etc., can solve the problems of high non-specific background, insufficient reaction, low degree of integration, etc., and achieve low cost , high sensitivity, simple operation effect
Active Publication Date: 2018-04-17
ZHEJIANG PUSHKANG BIOTECHNOLOGY CO LTD
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[0006] The biological microfluidic disk of immunomagnetic beads is an analysis and detection method that integrates magnetic particle technology and immunoassay on a microfluidic chip. At present, the main difficulties of this comprehensive detection method are as follows: For the intelligent control of internal micro-flow, the method commonly used at present is to set multiple micro-pumps and micro-valves inside the chip, which makes the micro-fluidic system more complicated; 2) Insufficient mixing of the reaction system leads to insufficient reaction; 3) The degree of integration is not high, resulting in high non-specific background
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Embodiment 1
[0022] Example 1: Quantitative detection of human chorionic gonadotropin β subunit
[0023] A chorionic gonadotropin beta subunit microfluidic disc of this embodiment, such as figure 1 , 2 As shown, it includes a microfluidic disc 13, and 12 microfluidic detection units are installed on the microfluidic disc 13, and each microfluidic detection unit includes a whole blood injection tank 1, and the whole blood injection The tank 1 is connected to the blood cell storage tank 3 through the whole blood separation channel 2, and the blood cell storage tank 3 is connected to the plasma delivery channel 4; the plasma delivery channel 4 is connected to the top of the mixing / detection tank 7; The top of the mixing / detection tank 7 communicates with the injection tank 9 of the magnetic particle solution coated with human chorionic gonadotropin beta subunit monoclonal antibody through a pipeline; the top of the mixing / detection tank 7 is also connected with the cleaning tank through a pi...
Embodiment 2
[0042] Example 2: Size screening of magnetic particles
[0043] The particle size of the magnetic microsphere is small, the specific surface area is large, and the surface contains active groups, so the coupling capacity is large, but the size of the magnetic microsphere is too small, which is not conducive to magnet collection, and the size is too large, which is not conducive to the mixing reaction in the disc. Magnetic particle size screening.
[0044] Refer to Example 1 for other experimental conditions, and the particle size of the magnetic particles is determined according to the following scheme.
[0045] Choose magnetic particle-labeled procalcitonin monoclonal antibodies with particle sizes of 0.1 μm, 0.5 μm, 1.0 μm, 1.5 μm, 2.0 μm, 3.0 μm, and 5.0 μm, respectively. The permanent magnet whose magnetic size has been optimized is used in the detection to fix the height of the magnet.
[0046] The experimental results are as follows:
[0047] The particle size of magn...
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Abstract
The invention discloses a chorionic gonadotropin beta subunit microfluidic disc and a use method thereof. The chorionic gonadotropin beta subunit microfluidic disc comprises a microfluidic disc body. A microfluidic detection unit is installed on the microfluidic disc body and comprises a whole blood injection groove, a whole blood separation channel, a blood cell storage groove, a plasma conveying channel, a mixing / detection groove, a magnetic particle solution injection groove, a cleaning fluid injection groove, an outer magnet area, a waste liquid tank, a human chorionic gonadotropin beta subunit monoclonal antibody solution injection groove and a light-emitting substrate solution injection groove, wherein the magnetic particle solution injection groove is wrapped by a human chorionic gonadotropin beta subunit monoclonal antibody, the outer magnet area contains a magnet, and the human chorionic gonadotropin beta subunit monoclonal antibody solution injection groove is labeled by alkaline phosphatase. By means of the simple method, intelligent control of microfluid in the microfluidic disc can be achieved. Meanwhile, reaction solutions can be fully mixed, it is guaranteed that a reaction system is efficient, and quantitative detection of the hCG concentration in samples is quickly achieved. The microfluidic disc has the advantages of being easy to operate, high in detection sensitivity, accurate and reliable in result, good in repeatability and low in cost.
Description
technical field [0001] The invention relates to a chorionic gonadotropin β subunit microfluidic disk and a method for using the same, belonging to the field of medical immunology in vitro diagnosis, capable of detecting the human chorionic gonadotropin β subunit in a biological sample in a very short time Quantitative detection has the characteristics of simple operation, high sensitivity and low cost. Background technique [0002] Human chorionic gonadotropin (Human Chorionic Gonadotropin, hCG) is a glycoprotein hormone synthesized and secreted by placental syncytiotrophoblast cells. It consists of two different subunits, α and β. Hormone), LH (luteinizing hormone) and TSH (thyroid-stimulating hormone) α-subunit structures are similar and can cross-react with each other. The β subunit is different from the β subunit of LH and has stronger specificity, so it becomes the preferred target in detection, and the total β-hCG can largely replace the indicator function of total hC...
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