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Magnetic particle chemiluminescence micro-fluidic chip

A microfluidic chip and chemiluminescent technology, applied in chemical instruments and methods, laboratory utensils, laboratory containers, etc., can solve problems such as incomplete reaction, stop of chip liquid flow, and inability of liquid to fill predetermined channels, etc. Achieve the effect of increasing accuracy and avoiding simple structural design

Inactive Publication Date: 2016-01-06
SHENZHEN HUAMAIXINGWEI MEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the actual situation is not necessarily ideal. Due to the chip or the sample, the reaction in the microfluidic chip may not proceed according to the set process. Filling each predetermined channel, etc., these will make the reaction between the reagents or substrates of the predetermined reaction incomplete, causing errors in the analysis results
However, since the reaction in the chip is done automatically, what the experimenter or detector sees is only the final result, and lacks the monitoring of the intermediate process, which will inevitably lead to wrong analysis of the result, thus affecting the accuracy of the analysis

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1: Microfluidic chip with liquid sensor device

[0041] like Figure 4 Shown is a schematic cross-sectional view of a liquid sensing device used in a microfluidic chip. A conductive needle (31) is inlaid on the microfluidic chip (27), and one end of the contact probe (30) is installed on the supporting circuit board (29), and the other end passes through the metal substrate (28) to connect with the microfluidic chip. contact with exposed conductive pins. The top of the metal substrate is used to carry the microfluidic chip and fix the circuit board, and the supporting circuit board is used to detect the electrical signal on the probe.

[0042] like Figure 5 and Figure 6 As shown, a plurality of conductive needles are inlaid on different positions of the microfluidic chip, all of which are located next to the microchannel through which the liquid passes through the microfluidic chip, and communicate with the microchannel. The conductive needle is made of a...

Embodiment 2

[0046] Example 2: Double-antibody sandwich determination of hypersensitive troponin I (cTnI)

[0047] (1) Antibody labeling

[0048] Dissolve 5 μg of HRP in 1 mL of distilled water, then add 0.2 mL of 0.1 M freshly prepared NaIO4 solution, react in the dark at room temperature for 20 min, and dialyze the purified solution with 1 mM pH 4.4 sodium acetate buffer. Then adjust the pH to 9.0 with 0.2 M pH9.5 carbonate buffer, add 10 μg of anti-cTnI monoclonal antibody, and react at room temperature for 2 h in the dark. Add 0.1 mL of newly prepared 4 mg / mL NaBH4 solution, mix well, and react at 4°C for 2 h. Put the above solution into a dialysis bag, dialyze against 0.15MpH7.4PBS, overnight at 4°C, and obtain HRP-labeled cTnI antibody.

[0049] Add 1mg magnetic particles (2μm in size), 10μg EDC and 15μg NHS solution and 15μg anti-cTnI monoclonal antibody (different from HRP-labeled antibody) solution to 1ml 10mM pH7.4 phosphate buffer, mix well and react at room temperature for 4h...

Embodiment 3

[0060] Example 3: Magnetic Particle Size Screening

[0061] Refer to Example 2 for other experimental conditions, and the magnetic particle size and magnetic induction of the magnet are carried out according to the following scheme.

[0062] The particle size is 0.1 μm, 0.5 μm, 1.8 μm, 2.4 μm, 1 μm, 3 μm, 10 μm. The magnetic induction of the magnet is 500 Gauss, 1000 Gauss, 4000 Gauss, 8000 Gauss, 12000 Gauss. Magnetic particles of seven sizes are driven by these five kinds of magnets respectively.

[0063] The experimental results show that when the 0.1μm magnetic particles are combined with the 500 Gauss magnet, the minimum detection limit is 500pg / ml, the quantitative detection range is 0.5~50ng / ml, and the linear correlation coefficient R 2 >0.95; intra-assay and inter-assay repeatability are less than 20%. That is: the chemiluminescent signal is weak, the sensitivity is not high, and the repeatability is poor.

[0064] When 10μm magnetic particles and 12000 Gauss magn...

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Abstract

The invention discloses a magnetic particle chemiluminescence micro-fluidic chip. The micro-fluidic chip comprises a top plate (1) and a bottom plate (2), the top plate (1) comprises an air pump (3), a sample adding opening (4), a sample filling area (12), a marking ligand storage pool (5) and a sample mixing area (13). The bottom plate (2) comprises a filter area (6), a magnetic particle coating area (7), a washing area (14), a detecting area (8), a washing liquid storage pool (9), a light-emitting substrate storage pool (10) and a liquid release channel (16). The top plate and the bottom plate both comprise liquid sensing devices, and the liquid sensing devices are used for determining the flowing state of liquid in the micro-fluidic chip and whether bubbles are mixed into the micro-fluidic chip.

Description

technical field [0001] The invention belongs to the technical field of microfluidic chip chemiluminescence immunoassay detection, and in particular relates to a chip which utilizes magnetic particle chemiluminescent technology and microfluidic chip technology to realize highly sensitive quantitative detection of analytes. Background technique [0002] At present, there are two main development trends for in vitro diagnostics (IVD): one is automation and integration, that is, the use of fully automated and highly sensitive large-scale instruments and equipment in the central laboratory of a large hospital to achieve high-precision disease analysis and diagnosis ; Another kind of miniaturization and bedside, that is, to realize rapid analysis and diagnosis on the spot through small and simple handheld devices. Small hospitals with insufficient funds and small sample sizes are not suitable for purchasing large and expensive equipment. At this stage, small rapid detection equip...

Claims

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Application Information

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IPC IPC(8): B01L3/00
Inventor 姜润华李泉
Owner SHENZHEN HUAMAIXINGWEI MEDICAL TECH CO LTD
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