CN-115634723-B - Microfluidic protein chip for allergen detection

CN115634723BCN 115634723 BCN115634723 BCN 115634723BCN-115634723-B

Abstract

The application belongs to the field of microfluidic protein chips, and particularly discloses a microfluidic protein chip for allergen detection, which comprises a sample adding area and a microfluidic unit, wherein the microfluidic unit comprises a dilution area, a premixing area, a marking area, a quantifying area and a detection area which are sequentially connected, the sample adding area is provided with a first sample adding area and a second sample adding area, the first sample adding area is used for adding whole blood samples, the second sample adding area is used for adding serum or plasma samples, different samples can be distinguished, the first sample adding area is used as a valve, the design is ingenious, a mixing ball is driven to vibrate by a vibrator in the premixing area, the mixing efficiency of the samples and the diluent is improved, the mixing time is reduced, the difference of detection results is reduced, the quantification of each sub-quantifying area is used for further reducing the difference of the detection results, and the marking area and the detection area can be coated with a plurality of antibodies, and meanwhile, the detection of a plurality of to-be-detected objects is carried out.

Inventors

TAO RUI
XIE HONGMEI
LI LIANGZHI
QIN FENGYAN
TANG TING

Assignees

苏州邦器生物技术有限公司

Dates

Publication Date: 20260508
Application Date: 20221017

Claims (7)

1. The microfluidic protein chip for allergen detection is characterized by comprising a sample adding area and at least one microfluidic unit, wherein the microfluidic unit comprises a dilution area, a premixing area, a marking area, a quantifying area and a detection area which are connected in sequence; The middle part of the microfluidic protein chip is a rotary installation part and is provided with a rotary center, and the rotary center is the rotary center during centrifugal operation; The sample adding area comprises a first sample adding area, a second sample adding area, a buffer area, a first waste liquid area, a separation area and a first capillary flow channel, wherein the first sample adding area and the second sample adding area are arranged around a rotation center, the first sample adding area is connected with the buffer area through a first separation front area and a first separation rear area, and the second sample adding area is directly connected with the buffer area; The first sample adding area and the second sample adding area are formed around the rotation center, and the first sample adding area is used for adding a whole blood sample; the first sample adding area is directly connected with the first separation front area, the first separation front area is far away from the rotation center relative to the first sample adding area, a filtering area is arranged between the first separation front area and the first separation rear area, a whole blood filtering membrane is arranged in the filtering area, whole blood is filtered by the whole blood filtering membrane through centrifugation, blood cells are filtered by the whole blood filtering membrane, plasma enters the first separation rear area, the first separation rear area is far away from the rotation center relative to the first separation front area, and the first separation rear area is connected with the buffer area through a first capillary flow channel; The first waste liquid zone is connected with the first separation front zone through an overflow flow channel; the buffer area is connected with the dilution area; A diluting solution is arranged in the diluting zone; The premixing zone is used for mixing a sample and a diluent, and comprises a first premixing zone and a second premixing zone, wherein the first premixing zone is internally provided with a first bulge, a second bulge, a third bulge, a fourth bulge, a fifth bulge, a mixing ball, a filter membrane, a vibrator and a first sensor; The first premixing zone is internally provided with a first partition, a second partition, a third partition and a fourth partition, the first partition, the second partition, the third partition and the fourth partition are separated by a filter membrane, the mixing balls freely move in the partitions, and the first bulge, the second bulge, the third bulge, the fourth bulge and the fifth bulge are fixedly arranged; a labeled antibody is arranged in the labeled region; The quantitative area is used for quantifying the mixed solution; the detection area is internally provided with a detection antibody.
2. The microfluidic protein chip for allergen detection according to claim 1, wherein the labeled antibody is at least one of allergens and the detection antibody is at least one of allergens.
3. The microfluidic protein chip for allergen detection according to claim 1, wherein a vibrator and a first sensor are arranged in the first premixing area, the main control module is connected with the vibrator and the first sensor, and the main control module controls the vibrator to work after receiving signals of the first sensor.
4. The microfluidic protein chip for allergen detection according to claim 1, wherein the first protrusion, the second protrusion, the third protrusion, the fourth protrusion, and the fifth protrusion have triangular cross-sectional shapes; The first bulge, the second bulge, the third bulge, the fourth bulge and the fifth bulge are symmetrically arranged about a vertical center line.
5. The microfluidic protein chip for allergen detection according to claim 1, wherein the quantification zone comprises a plurality of sub-quantification zones and a plurality of second waste liquid zones, wherein the second sensor is disposed in the second waste liquid zone.
6. The microfluidic protein chip for allergen detection according to claim 5, wherein a check valve is arranged at the joint of the sub-quantitative region and the detection region, the main control module is connected with the second sensor and the check valve, and the main control module controls the check valve to be opened or closed after receiving the signal of the second sensor.
7. The microfluidic protein chip for allergen detection according to claim 1, wherein the labeling region is provided with a quality control marker, and the detection region is coated with a quality control antibody, and the quality control antibody can specifically bind to the quality control marker.

Description

Microfluidic protein chip for allergen detection Technical Field The application belongs to the technical field of microfluidic protein chips, and relates to a microfluidic protein chip for allergen detection. Background The biochip is a new technical platform developed in the 90 th century of the 20 th century, and has the advantages of integration, high flux and high density. The microfluidic chip technology (Microfluidics) integrates basic operation units of sample preparation, reaction, separation, detection and the like in biological, chemical and medical analysis processes on a micron-scale chip, and automatically completes the whole analysis process. According to the different biomolecules fixed on the solid phase carrier, the microfluidic chip can be divided into a DNA chip, a protein chip and the like, and with the further deep research, the microfluidic protein chip is widely applied to a plurality of research fields, combines the characteristics of high flux, high sensitivity and high specificity of the microarray, and can conveniently, rapidly and accurately detect multiple samples and multiple items, and the microfluidic protein chip can process and analyze hundreds of samples within a few minutes. In a microfluidic protein chip, the fact that the difference between detection results of the same sample in different batches or the same sample in the same batch is large is common, because the viscosity of a blood sample is generally larger than that of a diluent, in a pipeline of the microfluidic protein chip, the important factors influencing the flow rate are capillary force and fluid viscosity, if the sample is not uniformly mixed with the diluent, the problem that the flow rate of a mixture of the sample and the diluent is inconsistent exists, and therefore the repeatability of the detection results is poor. Although the patent with publication number CN110026257B discloses a microfluidic chip, the purpose of uniformly mixing the sample and the diluent is achieved by arranging the filler in the premixing cavity, but there is still a problem of insufficient mixing. Disclosure of Invention The application aims to provide a microfluidic protein chip for allergen detection, which improves a premixing structure in the microfluidic protein chip, greatly improves the sufficiency of mixing a sample and a diluent, reduces the mixing time and reduces the residues of the sample and the diluent in a pipeline. In order to solve the technical problems, the application adopts the following technical scheme: According to one aspect of the application, the application provides a microfluidic protein chip for allergen detection, which comprises a sample adding area and at least one microfluidic unit, wherein the microfluidic unit comprises a dilution area, a premixing area, a marking area, a quantifying area and a detection cavity which are connected in sequence; The sample adding areas comprise at least two sample adding areas, wherein one sample adding area is used for adding a whole blood sample, and the other sample adding area is used for adding a serum or plasma sample; A diluting solution is arranged in the diluting zone; The premixing zone is used for mixing a sample and a diluent, and comprises a first premixing zone and a second premixing zone, wherein the first premixing zone is internally provided with a first bulge, a second bulge, a third bulge, a fourth bulge, a fifth bulge, a mixing ball, a filter membrane, a vibrator and a first sensor; a labeled antibody is arranged in the labeled region; The quantitative area is used for quantifying the mixed solution; the detection area is internally provided with a detection antibody. In one embodiment, the labeled antibody is at least one of an allergen and the detection antibody is at least one of an allergen. In one embodiment, a whole blood filter is disposed in the sample application region. In one embodiment, a first capillary flow passage is arranged in the sample adding region, one end of the first capillary flow passage is connected with the first separated region, and the other end of the first capillary flow passage is connected with the buffer region. In one embodiment, a first partition, a second partition, a third partition and a fourth partition are arranged in the first premixing area, and the first partition, the second partition, the third partition and the fourth partition are separated by a filter membrane; The mixing balls can freely move in each partition; The first bulge, the second bulge, the third bulge, the fourth bulge and the fifth bulge are fixedly arranged. In one embodiment, a vibrator and a first sensor are arranged in the first premixing area, a main control module is connected with the vibrator and the first sensor, and the main control module controls the vibrator to work after receiving signals of the first sensor. In one embodiment, the cross-sectional shapes of the first protrusion, the second protrusion, the third pro