CN116047059A - Biochip, kit and method for detecting target object to be detected - Google Patents
Biochip, kit and method for detecting target object to be detected Download PDFInfo
- Publication number
- CN116047059A CN116047059A CN202211732991.3A CN202211732991A CN116047059A CN 116047059 A CN116047059 A CN 116047059A CN 202211732991 A CN202211732991 A CN 202211732991A CN 116047059 A CN116047059 A CN 116047059A
- Authority
- CN
- China
- Prior art keywords
- liquid
- quality control
- sample
- antibody
- biochip
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 238000000018 DNA microarray Methods 0.000 title claims abstract description 117
- 238000000034 method Methods 0.000 title claims abstract description 31
- 239000007788 liquid Substances 0.000 claims abstract description 301
- 238000003908 quality control method Methods 0.000 claims abstract description 229
- 238000001514 detection method Methods 0.000 claims abstract description 124
- 239000012295 chemical reaction liquid Substances 0.000 claims abstract description 82
- 238000012360 testing method Methods 0.000 claims abstract description 79
- 238000012545 processing Methods 0.000 claims abstract description 54
- 238000002493 microarray Methods 0.000 claims abstract description 51
- 239000007790 solid phase Substances 0.000 claims abstract description 35
- 230000008569 process Effects 0.000 claims abstract description 15
- 229910052707 ruthenium Inorganic materials 0.000 claims description 45
- 238000006243 chemical reaction Methods 0.000 claims description 44
- 239000000758 substrate Substances 0.000 claims description 43
- 230000002159 abnormal effect Effects 0.000 claims description 30
- 239000012530 fluid Substances 0.000 claims description 29
- KJTLSVCANCCWHF-UHFFFAOYSA-N Ruthenium Chemical compound [Ru] KJTLSVCANCCWHF-UHFFFAOYSA-N 0.000 claims description 26
- 239000003550 marker Substances 0.000 claims description 23
- 210000002966 serum Anatomy 0.000 claims description 17
- 238000004020 luminiscence type Methods 0.000 claims description 15
- 239000012491 analyte Substances 0.000 claims description 8
- 238000002156 mixing Methods 0.000 claims description 7
- 239000012528 membrane Substances 0.000 claims description 6
- HWYHZTIRURJOHG-UHFFFAOYSA-N luminol Chemical compound O=C1NNC(=O)C2=C1C(N)=CC=C2 HWYHZTIRURJOHG-UHFFFAOYSA-N 0.000 claims description 5
- 210000002381 plasma Anatomy 0.000 claims description 5
- 210000003296 saliva Anatomy 0.000 claims description 5
- 239000007787 solid Substances 0.000 claims description 4
- 241000283707 Capra Species 0.000 claims description 3
- 239000004793 Polystyrene Substances 0.000 claims description 3
- 239000011521 glass Substances 0.000 claims description 3
- 239000004033 plastic Substances 0.000 claims description 3
- 229920002223 polystyrene Polymers 0.000 claims description 3
- 210000002700 urine Anatomy 0.000 claims description 3
- 208000023275 Autoimmune disease Diseases 0.000 claims description 2
- 201000006306 Cor pulmonale Diseases 0.000 claims description 2
- 206010028980 Neoplasm Diseases 0.000 claims description 2
- 239000003153 chemical reaction reagent Substances 0.000 abstract description 9
- 239000000523 sample Substances 0.000 description 218
- 239000000243 solution Substances 0.000 description 44
- 108090000790 Enzymes Proteins 0.000 description 24
- 102000004190 Enzymes Human genes 0.000 description 24
- 238000007789 sealing Methods 0.000 description 15
- 108090000623 proteins and genes Proteins 0.000 description 13
- 230000000903 blocking effect Effects 0.000 description 12
- 239000007853 buffer solution Substances 0.000 description 12
- 238000004140 cleaning Methods 0.000 description 12
- 102000004169 proteins and genes Human genes 0.000 description 11
- 239000000439 tumor marker Substances 0.000 description 6
- 238000009396 hybridization Methods 0.000 description 4
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 3
- 239000007993 MOPS buffer Substances 0.000 description 3
- 108010058846 Ovalbumin Proteins 0.000 description 3
- 239000007983 Tris buffer Substances 0.000 description 3
- 230000009471 action Effects 0.000 description 3
- 229940098773 bovine serum albumin Drugs 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 229940092253 ovalbumin Drugs 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 3
- 239000000427 antigen Substances 0.000 description 2
- 102000036639 antigens Human genes 0.000 description 2
- 108091007433 antigens Proteins 0.000 description 2
- 239000003085 diluting agent Substances 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000012634 fragment Substances 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 239000012488 sample solution Substances 0.000 description 2
- 230000008961 swelling Effects 0.000 description 2
- OOIBFPKQHULHSQ-UHFFFAOYSA-N (3-hydroxy-1-adamantyl) 2-methylprop-2-enoate Chemical compound C1C(C2)CC3CC2(O)CC1(OC(=O)C(=C)C)C3 OOIBFPKQHULHSQ-UHFFFAOYSA-N 0.000 description 1
- 102100023635 Alpha-fetoprotein Human genes 0.000 description 1
- 102100022785 Creatine kinase B-type Human genes 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 101001047117 Homo sapiens Creatine kinase B-type Proteins 0.000 description 1
- 101000998011 Homo sapiens Keratin, type I cytoskeletal 19 Proteins 0.000 description 1
- 102100033420 Keratin, type I cytoskeletal 19 Human genes 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 102100022019 Pregnancy-specific beta-1-glycoprotein 2 Human genes 0.000 description 1
- 102400001175 Unique cartilage matrix-associated protein C-terminal fragment Human genes 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 239000007822 coupling agent Substances 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 238000010166 immunofluorescence Methods 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 238000007781 pre-processing Methods 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 239000004065 semiconductor Substances 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54306—Solid-phase reaction mechanisms
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
- G01N33/581—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with enzyme label (including co-enzymes, co-factors, enzyme inhibitors or substrates)
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6854—Immunoglobulins
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Analytical Chemistry (AREA)
- Biotechnology (AREA)
- Pathology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Cell Biology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The application discloses biochip, biochip include solid phase carrier, are provided with liquid level control microarray on the solid phase carrier, and liquid level control microarray includes at least one in sample processing liquid control site, sample liquid control site, test reaction liquid control site and the luminous liquid control site. Aiming at the false negative problem possibly caused by inaccurate reagent sucking amount in the biochip detection process, the application creatively sets a plurality of liquid level quality control units on the surface of the solid phase carrier of the biochip, realizes liquid level quality control on the adding amount of different adding reagents, can quickly cause certain false negative problems, reduces unknown interference and reduces detection cost.
Description
Technical Field
The application relates to the technical field of biological detection, in particular to a biochip, a kit and a method for detecting a target object to be detected.
Background
Biochips, also known as protein chips or gene chips, originate from crystallization of DNA hybridization probe technology in combination with semiconductor industry technology. The technique refers to hybridization of a large number of probe molecules to fluorescent labeled DNA or other sample molecules (such as proteins, factors or small molecules) after immobilization on a support, and the number and sequence information of the sample molecules are obtained by detecting the hybridization signal intensity of each probe molecule.
At present, a biochip reading instrument is used in a process of sucking sample treatment liquid through a liquid-transferring gun, adding the sample treatment liquid into a chip cup, taking serum from a serum tube through the liquid-transferring gun, pushing the serum into the chip cup to react with the sample treatment liquid, placing a chip into the chip cup to react with a sample treatment liquid mixture, and then washing the chip to react with a reaction liquid and a luminous liquid. The operation is finished by automatic sample adding of a machine, and when the sample processing liquid, the sample liquid, the reaction liquid and the luminescent liquid are sucked, suction errors are easily generated, so that detection errors are caused by inaccurate suction amount of part of detection indexes.
Therefore, how to avoid the difficulty of detecting the biochip that the detection error is caused by inaccurate amount of the detection reagent.
Disclosure of Invention
In order to solve the above problems and avoid the interference caused by inaccurate detection reagent sucking amount, a first object of the present application is to provide a biochip, which includes a solid phase carrier, on which a liquid level quality control microarray is disposed, the liquid level quality control microarray includes at least one of a sample processing liquid quality control site, a sample liquid quality control site, a test reaction liquid quality control site, and a luminescence liquid quality control site; the sample treatment fluid comprises a ruthenium-containing substrate, and a first quality control antibody for specifically combining with the ruthenium-containing substrate is arranged at a quality control site of the sample treatment fluid; the sample liquid quality control site is provided with a second quality control antibody which is used for specifically combining with a non-target object to be detected in the sample liquid; the test reaction solution comprises a detection antibody which specifically binds to a target object to be detected, and a third quality control antibody which is used for specifically binding to the detection antibody is arranged at a quality control site of the test reaction solution; the luminescent liquid contains luminescent substrates for luminescent reaction with the first quality control antibody, the second quality control antibody and the detection antibody, at least one of a marker for luminescent reaction with the luminescent substrates and a fourth quality control antibody is arranged at a quality control site of the luminescent liquid, the fourth quality control antibody is not combined with a specific site of the detection antibody on a target object to be detected, the detection antibody and the fourth quality control antibody are both marked with markers, and the markers are used for luminescent reaction with the luminescent substrates.
Aiming at the false negative problem possibly caused by inaccurate reagent sucking amount in the biochip detection process, the application creatively sets a plurality of liquid level quality control units on the surface of the solid phase carrier of the biochip, realizes liquid level quality control on the adding amount of different adding reagents, can quickly cause certain false negative problems, reduces unknown interference and reduces detection cost.
In one embodiment, the liquid level quality control microarray quality control site satisfies at least one of the following features (1) - (6):
(1) The luminescent liquid comprises luminol and H 2 O 2 ；
(2) The marker is HRP;
(3) The sample liquid is selected from at least one of plasma, serum, saliva and urine;
(4) The first quality control antibody arranged at the quality control site of the sample treatment fluid is an anti-ruthenium antibody, and the anti-ruthenium antibody is used for specifically combining with a ruthenium-containing substrate in the sample treatment fluid;
(5) The second quality control antibody arranged at the sample liquid quality control site is an anti-human IgG antibody;
(6) And the third quality control antibody arranged at the quality control site of the test reaction liquid is an anti-marker antibody.
In one embodiment, the solid support meets at least one of the following features (1) - (2):
(1) The solid phase carrier is selected from at least one of a black glass slide, a polystyrene plastic carrier, an NC membrane carrier and a PDVF membrane carrier;
(2) The solid phase carrier is also provided with at least one of a positive quality control microarray, a negative quality control microarray and a test microarray for detecting a target object to be detected.
A second object of the present application is to provide a kit comprising the above biochip.
In one embodiment, the sample processing device further comprises at least one of a sample processing liquid, a sample quality control reaction liquid, a test reaction liquid and a luminescence liquid.
In one embodiment, the kit satisfies at least one of the following features (1) to (3):
(1) The sample treatment fluid comprises a ruthenium-containing substrate;
(2) The sample quality control reaction liquid comprises an anti-human IgG antibody, and the anti-human IgG antibody in the sample quality control reaction liquid is marked with a marker;
(3) The test reaction liquid comprises a detection antibody and an anti-ruthenium antibody, and the anti-ruthenium antibody in the test reaction liquid is marked with a marker;
(4) The luminous liquid comprises luminol and H 2 O 2 。
In one embodiment, the kit satisfies at least one of the following features (1) to (3):
(1) The content of the substrate in the sample treatment liquid is 5-20 mug/mL;
(2) The anti-human IgG antibodies include at least one of goat anti-human IgG antibodies and mouse anti-human IgG antibodies;
(3) The anti-human IgG antibody in the sample quality control reaction solution is marked with a marker.
A third object of the present application is to provide a method for manufacturing a biochip, comprising the steps of:
setting a sample treatment liquid quality control site, a sample liquid quality control site, a reaction liquid quality control site and a luminescence liquid quality control site on a solid phase carrier to obtain a liquid level quality control microarray;
and (3) sealing the solid phase carrier provided with the liquid level quality control microarray by using a sealing liquid to prepare the biochip.
In one embodiment, the steps of the method for producing a biochip satisfy at least one of the following features (1) to (3):
(1) The liquid level quality control microarray is arranged in a sample application mode;
(2) The blocking of the solid phase carrier with the blocking liquid specifically comprises:
immersing the solid phase carrier provided with the liquid level quality control microarray into the sealing liquid for 1-4 hours, taking out the sealed solid phase carrier, and removing residual sealing liquid to prepare a biochip;
(3) The blocking solution is a buffer solution containing blocking proteins.
In one embodiment, the sealing liquid satisfies at least one of the following characteristics (1) to (2):
(1) The blocking protein is selected from at least one of bovine serum albumin and ovalbumin;
(2) The buffer solution is at least one selected from PBS buffer solution, tris buffer solution, HEPS buffer solution and MOPS buffer solution.
A fourth object of the present application is to provide a method for detecting a target analyte, the method comprising:
detecting a target object to be detected in the sample liquid by adopting the kit;
determining whether the addition amount of at least one of the sample processing liquid, the sample quality control reaction liquid, the test reaction liquid and the luminescent liquid is abnormal in the detection process according to the detection signals of the liquid level quality control microarray of the biochip in the kit.
In one embodiment, detecting a target object in a sample solution by using the kit specifically includes:
mixing a sample liquid with a sample processing liquid in a kit to obtain a sample mixed liquid, reacting the sample mixed liquid with a biochip in the kit, and reacting the reacted biochip with a test reaction liquid and a luminescent liquid;
and/or
Mixing the sample liquid and the sample processing liquid to obtain a sample mixed liquid, reacting the sample mixed liquid with the biochip in the kit, and reacting the reacted biochip with the sample quality control reaction liquid and the luminescent liquid in the kit.
In one embodiment, determining whether the addition amount of at least one of the sample processing liquid, the sample quality control reaction liquid, the test reaction liquid and the luminescence liquid is abnormal in the detection process according to the detection signal of the quality control site of the biochip in the kit specifically includes:
determining whether the addition amount of the corresponding test reaction liquid and/or luminous liquid is abnormal or not according to the detection signals of the test reaction liquid quality control site and/or luminous liquid quality control site of the biochip;
and/or
If the sample quality control reaction liquid is adopted to react with the biochip, determining whether the addition amount of the sample liquid is abnormal or not according to the detection signal of the sample liquid quality control site of the biochip;
and/or
If the sample treatment liquid comprises a ruthenium-containing substrate, determining whether the addition amount of the sample liquid is abnormal according to the detection signal of the quality control site of the sample treatment liquid of the biochip.
In one embodiment, the sample fluid is selected from at least one of plasma, serum, saliva, and urine.
In one embodiment, the target analyte is selected from any one of a tumor marker, an autoimmune disease marker, and a cardiopulmonary disease marker.
Drawings
In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings that are needed in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present application, and other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a schematic diagram showing the structure of a biochip prepared in example 1 of the present application;
fig. 2 is a quality control result of the addition amount of the sample processing liquid in example 3 of the present application, in fig. 2, a represents a detection result of the addition amount of the sample processing liquid being sufficient, and b represents a detection result of the addition amount of the sample processing liquid being insufficient or not being sufficient;
fig. 3 is a quality control result of the addition amount of the sample liquid in example 4 of the present application, in fig. 3, a represents a detection result of the addition amount of the sample liquid being sufficient, and b represents a detection result of the addition amount of the sample liquid being insufficient or not being sufficient;
fig. 4 is a quality control result of the addition amount of the luminescent liquid in example 5 of the present application, in fig. 4, a represents a detection result of the addition amount of the luminescent liquid being sufficient, and b represents a detection result of the addition amount of the luminescent liquid being insufficient or not being sufficient;
fig. 5 shows the quality control result of the amount of the test reaction liquid in example 6 of the present application, where a shows the detection result of the addition of the test reaction liquid being sufficient, and b shows the detection result of the addition of the test reaction liquid being insufficient.
Detailed Description
Reference now will be made in detail to the embodiments of the application, one or more examples of which are described below. Each example is provided by way of explanation, not limitation, of the present application. Indeed, it will be apparent to those skilled in the art that various modifications and variations can be made to the present application without departing from the scope or spirit of the present application. For example, features illustrated or described as part of one embodiment can be used on another embodiment to yield still a further embodiment.
Accordingly, it is intended that the present application cover such modifications and variations as fall within the scope of the appended claims and their equivalents. Other objects, features and aspects of the present application are disclosed in or are apparent from the following detailed description. It is to be understood by one of ordinary skill in the art that the present discussion is a description of exemplary embodiments only, and is not intended as limiting the broader aspects of the present application.
The surface of the carrier of the biochip is generally provided with a negative quality control microarray, a positive quality control microarray and a microarray for detecting each target object to be detected, which are used for positioning chip data, whether HAMA interference influence exists or not and detecting each index. In the biochip detection process, false negative detection results may be caused by inaccurate suction amount in the operation process. However, the current biochip cannot analyze and determine the specific cause of the above false negatives.
In order to solve the above technical problems, a first aspect of the present application provides a biochip, the biochip includes a solid phase carrier, on which a liquid level quality control microarray is disposed, the liquid level quality control microarray includes at least one of a sample processing liquid quality control site, a sample liquid quality control site, a test reaction liquid quality control site, and a luminescence liquid quality control site; the sample treatment fluid comprises a ruthenium-containing substrate, and a first quality control antibody for specifically combining with the ruthenium-containing substrate is arranged at a quality control site of the sample treatment fluid; the sample liquid quality control site is provided with a second quality control antibody which is used for specifically combining with a non-target object to be detected in the sample liquid; the test reaction solution comprises a detection antibody which specifically binds to a target object to be detected, and a third quality control antibody which is used for specifically binding to the detection antibody is arranged at a quality control site of the test reaction solution; the luminescent liquid contains luminescent substrates for luminescent reaction with the first quality control antibody, the second quality control antibody and the detection antibody, at least one of a marker for luminescent reaction with the luminescent substrates and a fourth quality control antibody is arranged at a quality control site of the luminescent liquid, the fourth quality control antibody is not combined with a specific site of the detection antibody on a target object to be detected, the detection antibody and the fourth quality control antibody are both marked with markers, and the markers are used for luminescent reaction with the luminescent substrates.
As used herein, the term "biochip" generally refers to a microarray hybridization chip (micro-array) of bioinformatic molecules (e.g., gene fragments, DNA fragments or polypeptides, proteins, sugar molecules, tissues, etc.) immobilized on a supporting medium at high density, the sequence and position of each molecule in the array being known and being a predetermined array of sequences.
The detection of the biochip based on immunofluorescence requires the addition of a sample processing liquid, a test reaction liquid and a luminescent liquid in the detection process to detect the target object to be detected in the sample liquid. Generally, the source of the sample fluid is different according to the target object to be detected by the biochip. The target analyte to be detected by the biochip may be one or more. Specifically, the target to be detected by the biochip may be a tumor marker or other proteins, such as an IgG antibody, and further an IgG antibody may be a human IgG antibody or other animal IgG antibodies.
Specifically, the sample liquid is at least one selected from the group consisting of plasma, serum, and saliva. For the target test substance to be human IgG antibodies, the sample is derived from human accordingly.
The sample treatment liquid is used for preprocessing the sample liquid so as to reduce interference of other components in the sample on detection of the target object to be detected. The test reaction liquid contains detection antibodies, is used for specifically combining with target objects to be detected combined on the biochip in the detection process, and the luminous liquid is used for reacting with various quality control antibodies and the detection antibodies to generate detection signals, and the concentration of the target objects to be detected or the quality control of the addition amounts of the sample processing liquid, the sample liquid, the test reaction liquid and the luminous liquid can be determined according to the detection signals.
Aiming at the false negative problem possibly caused by inaccurate reagent sucking amount in the biochip detection process, the application creatively sets a plurality of liquid level quality control sites on the surface of the solid phase carrier of the biochip, realizes liquid level quality control on the adding amount of different adding reagents, can quickly cause certain false negative problems, reduces the influence of unknown interference and reduces the detection cost.
The sample treatment liquid quality control site is used for judging whether the addition amount of the sample treatment liquid is abnormal in the detection process of the target object to be detected; the sample liquid quality control site is used for judging whether the addition amount of the sample liquid is abnormal in the detection process of the target object to be detected; the test reaction liquid quality control site is used for judging whether the addition amount of the test reaction liquid is abnormal in the detection process of the target object to be detected; the luminous liquid quality control site is used for judging whether the adding amount of the luminous liquid is abnormal in the detection process of the target object to be detected.
In some embodiments, the first luminescent substrate in the sample processing liquid comprises a ruthenium-containing substrate, the first quality control antibody arranged at the quality control site of the sample processing liquid is an anti-ruthenium antibody, ruthenium is a catalytic coupling agent, the ruthenium-containing substrate can form a compound with the anti-ruthenium antibody, adding quantitative ruthenium-containing substrate in the sample processing liquid can not affect the product performance, capturing the added quantitative ruthenium-containing substrate in the sample processing liquid by using the anti-ruthenium antibody at the quality control site of the sample processing liquid, further reacting the captured ruthenium-containing substrate with the anti-ruthenium antibody marked by the marker additionally added in the test reaction liquid, and generating a detectable signal under the action of the luminescent liquid for judging whether the adding amount of the sample processing liquid is abnormal.
For the biochip for detecting the target object to be detected in the human blood sample or the serum sample, in some specific embodiments, the second quality control antibody arranged at the quality control site of the sample liquid is an anti-human IgG antibody, and the anti-human IgG antibody is specifically combined with the IgG antibody in the sample liquid to fix the IgG antibody in the sample at the quality control site of the sample, so that the second quality control antibody is specifically combined with the anti-human IgG antibody marked by the marker in the quality control reaction liquid of the sample to generate a detectable signal under the action of the luminescent liquid, so as to detect whether the addition amount of the sample liquid is abnormal. Accordingly, the sample quality control reaction solution contains an anti-human IgG antibody, and the anti-human IgG antibody in the sample quality control reaction solution is labeled with a label.
In some embodiments, the labeled antibody in the test reaction solution comprises a detection antibody; the detection antibody in the test reaction solution is labeled with a label. Correspondingly, a third quality control antibody arranged at the quality control site of the test reaction liquid is an anti-marker antibody so as to be specifically combined with the detection antibody, and a detectable signal is generated after the detection antibody reacts with the luminous liquid, so that the adding amount of the test reaction liquid is quality controlled. Correspondingly, the test reaction liquid also contains an anti-ruthenium antibody, and the anti-ruthenium antibody is marked with a marker to react with a ruthenium-containing substrate captured in the sample processing liquid and generate a detectable signal under the action of the luminescent liquid. In some embodiments, the luminescent substrate in the luminescent solution comprises luminol and H 2 O 2 . Correspondingly, the light-emitting liquid quality control site is provided with a marker or a fourth quality control antibody, the fourth quality control antibody is marked with the marker, and specifically, the fourth quality control antibody antigen is a specific antibody of a non-target object to be detected in the sample.
In some embodiments, the label is an HRP enzyme for use in combination with a luminescent substrate to produce a detectable signal. It is to be understood that the choice of the label and luminescent substrate of the present application is not limited to the above-mentioned limitations, as long as the combination of the label and luminescent substrate that does not affect the detection reaction of the present application and that can generate a detectable signal can be used for detection and liquid level quality control of the target analyte of the present application. Accordingly, a second aspect of the present application also provides a kit comprising the above biochip.
In some embodiments, the kit further comprises at least one of the sample processing fluid, the sample quality control reaction fluid, the test reaction fluid, and the luminescence fluid described above.
In some embodiments, the sample processing fluid includes a ruthenium-containing substrate for chemically reacting with a sample processing fluid quality control site.
In some embodiments, the sample-based control reaction solution comprises a labeled anti-human IgG antibody. More specifically, the concentration of the anti-human IgG antibody in the sample quality control reaction solution is 0.05-0.5. Mu.g/mL, further 0.1-0.5. Mu.g/mL, and still further 0.3-0.5. Mu.g/mL.
In some embodiments, the anti-human IgG antibodies comprise at least one of goat anti-human IgG antibodies and mouse anti-human IgG antibodies.
In some embodiments, the test reaction solution comprises a detection antibody and an anti-ruthenium antibody, each of which is labeled with a label that produces a detectable signal with the luminescent substrate. For detecting macromolecular antigens by a double-antibody sandwich method, the detection antibody can be a labeled primary antibody which specifically binds to other binding sites of the target object to be detected, the other binding sites refer to epitopes on the surface of the target object to be detected except for binding sites of the solid-phase carrier coated antibody, and for detecting the target object to be detected by using an indirect ELISA, the detection antibody in the test reaction solution can also be a labeled secondary antibody which specifically binds to the target object to be detected. In some embodiments, the detection antibody is a second enzyme-labeled antibody. More specifically, the enzyme-labeled secondary antibody is an HPR-labeled secondary antibody.
In some embodiments, the concentration of the detection antibody in the test reaction solution may be 0.1 to 2. Mu.g/mL, more preferably 0.1 to 1. Mu.g/mL, still more preferably 0.5 to 2. Mu.g/mL, and still more preferably 0.5 to 1. Mu.g/mL.
In some embodiments, the concentration of the anti-ruthenium antibody in the test reaction solution is 0.5 to 2. Mu.g/mL, more preferably 1 to 2. Mu.g/mL, still more preferably 1.5 to 2. Mu.g/mL.
In some embodiments, the substrate is present in the sample treatment fluid in an amount of 5 to 20 μg/mL. Further, the concentration may be 10 to 20. Mu.g/mL. Further, the concentration may be 15 to 20. Mu.g/mL.
In some embodiments, the solid support comprises at least one of a black slide, a polystyrene plastic support, an NC membrane support, and a PDVF membrane support.
In some embodiments, the solid support further has disposed thereon at least one of a positive quality control microarray, a negative quality control microarray, and a test microarray for detecting a target analyte. The target test substance may be various, for example, various tumor markers, and the corresponding biomolecules specifically binding to different target test substances may also include various, specifically, antibodies.
The second aspect of the present application provides a method for preparing the above kit, comprising the steps of:
corresponding antibodies are arranged on the sample treatment liquid quality control site, the sample liquid quality control site, the reaction liquid quality control site and the luminous liquid quality control site on the solid phase carrier to form a liquid level quality control microarray; the sample treatment liquid quality control site is provided with a first quality control antibody which is used for specifically combining with the ruthenium-containing substrate; the sample liquid quality control site is provided with a second quality control antibody which is used for specifically combining with a non-target object to be detected in the sample liquid; the test reaction liquid quality control site is provided with a third quality control antibody which is used for being specifically combined with the detection antibody; the luminous liquid quality control site is provided with at least one of a marker for carrying out luminous reaction with a luminous substrate and a fourth quality control antibody, the fourth quality control antibody is not combined with the specific site of the detection antibody on the target object to be detected, and the detection antibody and the fourth quality control antibody are both marked with markers.
And (3) sealing the solid phase carrier provided with the liquid level quality control microarray by using a sealing liquid to prepare the biochip.
Specifically, the blocking solution is a buffer solution containing blocking proteins and is used for blocking antibodies arranged on the liquid level quality control microarray; wherein the blocking protein is bovine serum albumin or ovalbumin; the buffer solution is at least one selected from PBS buffer solution, tris buffer solution, HEPS buffer solution and MOPS buffer solution.
In some embodiments, blocking the solid phase carrier provided with the liquid level control column with a blocking liquid comprises:
immersing the solid phase carrier provided with the liquid level quality control microarray into the sealing liquid for 1-4 h, taking out the solid phase carrier after sealing, and removing residual sealing liquid to obtain the biochip. Specifically, the blocking liquid may be removed by centrifugation.
In some embodiments, the liquid level quality control microarray is arranged in a spotting arrangement.
A third aspect of the present application provides a liquid level quality control method, including:
detecting a target object to be detected in the sample liquid by adopting the kit;
and determining whether the addition amount of at least one of the sample processing liquid, the sample liquid, the reaction liquid and the luminescent liquid is abnormal according to the detection signal of the quality control site of the biochip.
In some embodiments, the method comprises:
mixing a sample liquid with a sample treatment liquid in the kit to obtain a sample mixed liquid, reacting the sample mixed liquid with the biochip in the kit, and reacting the reacted biochip with a test reaction liquid and a luminescent liquid after treatment;
determining whether the addition amount of the test reaction liquid is abnormal according to the detection signal of the quality control site of the test reaction liquid of the biochip;
and/or
And determining whether the adding amount of the luminescent liquid is abnormal or not according to the detection signal of the luminescent liquid quality control site of the biochip.
In some embodiments, to detect whether the sample processing fluid addition is abnormal, the method includes:
mixing a sample liquid with a sample treatment liquid in the kit to obtain a sample mixed liquid, reacting the sample mixed liquid with the biochip in the kit, and reacting the reacted biochip with a test reaction liquid and a luminescent liquid after treatment;
and determining whether the addition amount of the sample processing liquid is abnormal according to the detection signal of the quality control site of the sample processing liquid of the biochip.
In other embodiments, in order to detect whether the addition amount of the sample liquid is abnormal, mixing the sample liquid and the sample treatment liquid to obtain a sample mixed liquid, reacting the sample mixed liquid with the biochip in the kit, and then treating the reacted biochip to react with the sample quality control reaction liquid and the luminescent liquid in the kit;
whether the addition amount of the sample liquid is abnormal or not is determined according to the detection signal of the quality control site of the sample processing liquid of the biochip, and meanwhile, whether the addition amount of the luminescent liquid is abnormal or not can also be determined.
In other embodiments, in order to detect whether the addition amounts of the sample liquid and the sample processing liquid are abnormal at the same time, the sample liquid and the sample processing liquid are mixed to obtain a sample mixed liquid, the sample mixed liquid is reacted with the biochip in the kit, and then the reacted biochip is treated and then reacted with the sample quality control reaction liquid and the luminescent liquid in the kit, wherein the sample processing liquid comprises a ruthenium-containing substrate;
determining whether the addition amount of the sample processing liquid is abnormal according to the detection signal of the quality control site of the sample processing liquid of the biochip; and
and determining whether the addition amount of the sample liquid is abnormal according to the detection signal of the sample liquid quality control site of the biochip.
It can be understood that, when detecting whether the addition amounts of the test sample processing liquid and the sample liquid are abnormal, whether the addition amount of the test reaction liquid is abnormal can be determined according to the detection signal of the quality control site of the test reaction liquid of the biochip, and whether the addition amount of the luminescent liquid is abnormal can be determined according to the detection signal of the quality control site of the luminescent liquid.
Embodiments of the present application will be described in detail below with reference to examples, but the present application is not limited to these examples. The test methods used in the following examples are conventional methods unless otherwise specified; the materials, reagents and the like used, unless otherwise specified, are those commercially available.
The embodiment of the application provides application of the biochip capable of performing liquid level quality control in tumor marker detection, and provides a chip for tumor marker detection, a kit for tumor marker detection and a liquid level quality control method for tumor marker detection. It is understood that the biochip, kit and liquid level quality control method of the present application are not limited to the swelling mark detection system, but are also applicable to the detection of other systems such as heart and lung, self-care, etc.
Example 1
The embodiment provides a biochip, as shown in fig. 1, the biochip comprises a solid phase carrier, the solid phase carrier is a black glass slide, a liquid level quality control microarray, a test microarray, a positive quality control microarray and a negative quality control microarray are arranged on the solid phase carrier, wherein the positive quality control microarray is arranged in an A region of the solid phase carrier, the test microarray is arranged in a B region of the solid phase carrier, the negative quality control microarray is arranged in a C region of the solid phase carrier, the liquid level quality control microarray is arranged in a D region of the solid phase carrier, wherein the test microarray is provided with a detection index antibody, and the detection index antibody comprises at least one of an AFP antibody, a CA-199 antibody, a CEA antibody, a PGI antibody, a PGII antibody, a GRP antibody, a CA-724 antibody, an HCG antibody, a CA-125 antibody, a CA-153 antibody, an NSE antibody and a CK19 antibody, the positive quality control microarray is provided with an anti-HRP antibody, the negative quality control microarray is provided with a negative quality control site, and the negative quality control site is provided with an antibody of a non-swelling chip detection system, for example: the CKBB antibody and liquid level quality control microarray comprises a sample treatment liquid quality control site 21, a sample liquid quality control site 21, a test reaction liquid quality control site 23 and a luminous liquid quality control site 24. The sample treatment liquid quality control site 21 is provided with an anti-ruthenium antibody; the sample liquid quality control site 22 is provided with an anti-human IgG antibody, wherein the anti-human IgG antibody can be sheep anti-human IgG or mouse anti-human IgG; the test reaction liquid quality control site 23 is provided with HRP enzyme; the luminescence control site 24 is provided with an anti-HRP antibody.
The preparation method of the biochip of the embodiment comprises the steps of:
the method comprises the steps that antibody sample points corresponding to a sample treatment liquid quality control site, a sample liquid quality control site, a reaction liquid quality control site and a luminescence liquid quality control site are arranged on a solid phase carrier to form a liquid level quality control microarray, and the solid phase carrier is provided with a test microarray, a positive quality control microarray and a negative quality control microarray;
immersing the solid phase carrier provided with the liquid level quality control microarray into a sealing solution for 1-4 h, taking out the solid phase carrier, centrifuging to remove residual sealing solution, and obtaining the biochip sealing solution which is a buffer solution containing sealing protein, wherein the sealing protein is bovine serum albumin or ovalbumin, and the buffer solution is any one of PBS buffer solution, tris buffer solution, HEPS buffer solution and MOPS buffer solution.
Wherein, the components such as the anti-ruthenium antibody, the anti-human IgG antibody, the HRP enzyme, the anti-HRP antibody and the like are purchased from Jinlian organisms, and the product numbers are shown in the following table 1.
TABLE 1
Example 2
The present embodiment provides a kit comprising the biochip of embodiment 1, a sample processing liquid, a sample quality control reaction liquid, a test reaction liquid, and a luminescent liquid. The sample treatment solution comprises a ruthenium-containing substrate, interference elimination protein Mak33 and partial index antibodies, such as PGI antibodies; the sample quality control reaction solution comprises an anti-human IgG antibody of a target enzyme; the test reaction solution comprises an enzyme-labeled secondary antibody. The content of the ruthenium-containing substrate in the sample treatment solution is 5-20 mug/mL; wherein, the ruthenium-containing substrate is purchased from Jinlian organism, the product number is McAb20190614, the luminescent liquid is purchased from Jinlian organism, and the product number is LRZ2018027.
Wherein the step of preparing a sample processing liquid comprises:
adding 1% -10% of Mak33, 5-20 mug/mL of ruthenium-containing substrate and 10-20 mug/mL of partial index antibody into ultrapure water, and adding 5-10 mug/mL of PGI antibody and 15-20 mug/mL of CA153 antibody to obtain sample treatment liquid;
the preparation method of the test reaction liquid and the sample quality control reaction liquid comprises the following steps:
the concentrations of each index antibody and the anti-ruthenium antibody were labeled with enzyme and diluted with a stable diluent (purchased from Jinlian, cat No. LRZ 2022025) according to the following table 2 to obtain test reaction solutions, and the anti-human IgG antibody was labeled with enzyme and diluted with the above diluent according to the concentrations of table 2 to prepare sample quality control reaction solutions.
TABLE 2
Example 3 detection of the amount of sample treatment solution added
The embodiment provides a liquid level quality control method, which utilizes the kit of embodiment 2 to control the liquid level quality. The liquid level quality control method of the embodiment specifically comprises the following steps: taking 20ul of sample treatment liquid by a liquid-transferring gun, adding the sample treatment liquid into a chip cup, taking 180ul of human serum sample liquid from a serum tube by the liquid-transferring gun, pushing the sample liquid into the chip cup to react with the sample treatment liquid for 40 minutes to obtain sample mixed liquid, placing the biochip in the kit in the example 2 into the chip cup to react with the sample mixed liquid, cleaning the residual sample mixed liquid on the surface of the biochip, then absorbing 200ul of enzyme-labeled secondary antibody reaction liquid, and cleaning the residual enzyme-labeled secondary antibody reaction liquid on the surface of the biochip after reacting with the biochip cleaned with the sample mixed liquid for 40 minutes, and absorbing 200ul of luminescent liquid and cleaning the biochip cleaned with the enzyme-labeled secondary antibody reaction liquid to perform luminescent reaction;
and acquiring detection signals of the quality control sites of the sample processing liquid of the biochip by using a biochip reading instrument, and determining whether the addition amount of the sample processing liquid is abnormal or not according to the detection signals of the quality control sites of the sample processing liquid of the biochip.
Specifically, as shown in fig. 2, a represents a detection result obtained by adding a sufficient amount of the sample processing liquid, and b represents a detection result obtained by not adding a sufficient amount of the sample processing liquid or not adding a sufficient amount of the sample processing liquid to the detection signal of the biochip. As can be seen from fig. 2, when the addition amount of the sample processing liquid is not sufficient or is not sufficient, the detection signal of the quality control site of the sample processing liquid is significantly lower than the detection signal of the addition amount of the sample processing liquid.
Example 4 detection of the amount of sample liquid added
The embodiment provides a liquid level quality control method, which utilizes the kit of embodiment 2 to control the liquid level quality. The liquid level quality control method of the embodiment specifically comprises the following steps: taking 20ul of sample treatment liquid by a liquid-transferring gun, adding the sample treatment liquid into a chip cup, taking 180ul of human serum sample liquid from a serum tube by the liquid-transferring gun, pushing the sample liquid into the chip cup to react with the sample treatment liquid for 40 minutes to obtain sample mixed liquid, placing the biochip in the kit in the example 2 into the chip cup to react with the sample mixed liquid, cleaning the residual sample mixed liquid on the surface of the biochip, then absorbing 200ul of enzyme-labeled secondary antibody reaction liquid, and cleaning the residual enzyme-labeled secondary antibody reaction liquid on the surface of the biochip after the biochip reacts with the cleaned biochip for 40 minutes, and absorbing 200ul of luminescent liquid and cleaning the biochip of the enzyme-labeled secondary antibody reaction liquid to react;
and acquiring detection signals of the sample liquid quality control sites of the biochip by using a biochip reading instrument, and determining whether the addition amount of the sample liquid is abnormal or not according to the detection signals of the sample liquid quality control sites of the biochip.
Specifically, as shown in fig. 3, the detection signal of the biochip is shown in fig. 3, a represents a detection result obtained by adding a sufficient amount of the sample liquid, and b represents a detection result obtained by not adding a sufficient amount of the sample liquid or not adding a sufficient amount of the sample liquid. As can be seen from fig. 3, when the amount of the sample liquid is not added or is not added, the detection signal of the sample liquid quality control site is significantly lower than the detection signal added by the amount of the sample liquid.
Example 5 detection of the amount of luminescent liquid to be added
The embodiment provides a liquid level quality control method, which comprises the following steps: taking 20ul of sample treatment liquid by a liquid-transferring gun, adding the sample treatment liquid into a chip cup, taking 180ul of human serum sample liquid from a serum tube by the liquid-transferring gun, pushing the sample treatment liquid into the chip cup to react with the sample treatment liquid to obtain a sample mixed liquid, placing the biochip in the kit in the example 2 into the chip cup to react with the sample mixed liquid, cleaning the residual sample mixed liquid on the surface of the biochip, then absorbing 200ul of enzyme-labeled secondary antibody reaction liquid, and cleaning the residual enzyme-labeled secondary antibody reaction liquid on the surface of the biochip after the biochip reacts with the cleaned biochip in 40 minutes, and absorbing 200ul of luminescent liquid and cleaning the biochip of the enzyme-labeled secondary antibody reaction liquid to react;
and acquiring detection signals of the quality control sites of the test reaction liquid of the biochip by using a biochip reading instrument, and determining whether the addition amount of the test reaction liquid is abnormal or not according to the detection signals of the quality control sites of the test reaction liquid of the biochip.
Specifically, as shown in fig. 4, the detection signal of the biochip is shown in fig. 4, a indicates a detection result that the addition amount of the luminescent liquid is sufficient, and b indicates a detection result that the addition amount of the luminescent liquid is not sufficient or is not sufficient. As can be seen from fig. 4, when the addition amount of the luminescence liquid is not sufficient or is not sufficient, the detection signals of the sample processing liquid quality control site, the sample liquid quality control site, the luminescence liquid quality control site, and the reaction liquid quality control site are all significantly lower than the detection signals obtained by adding the addition amount of the luminescence liquid.
Example 6 detection of the amount of added reaction solution
The embodiment provides a liquid level quality control method, which comprises the following steps: taking 20ul of sample treatment liquid by a liquid-transferring gun, adding the sample treatment liquid into a chip cup, taking 180ul of human serum sample liquid from a serum tube by the liquid-transferring gun, pushing the sample treatment liquid into the chip cup to react with the sample treatment liquid to obtain a sample mixed liquid, placing the biochip in the kit in the example 2 into the chip cup to react with the sample mixed liquid, cleaning the residual sample mixed liquid on the surface of the biochip, then absorbing 200ul of enzyme-labeled secondary antibody reaction liquid, and cleaning the residual enzyme-labeled secondary antibody reaction liquid on the surface of the biochip after the biochip reacts with the cleaned biochip in 40 minutes, and absorbing 200ul of luminescent liquid and cleaning the biochip of the enzyme-labeled secondary antibody reaction liquid to react;
and acquiring detection signals of the quality control sites of the test reaction liquid of the biochip by using a biochip reading instrument, and determining whether the addition amount of the test reaction liquid is abnormal or not according to the detection signals of the quality control sites of the test reaction liquid of the biochip.
Specifically, as shown in fig. 5, the detection signal of the biochip is shown in fig. 5, a indicates a detection result that the amount of the test reaction solution added is sufficient, and b indicates a detection result that the amount of the test reaction solution added is not sufficient. As can be seen from fig. 5b, when the amount of the added test reaction solution is insufficient, HRP enzyme in the dried test reaction solution is attached to the surface of the chip due to the long reaction time, so that the HRP enzyme is not washed clean when the chip is washed, and the uncleaned HRP enzyme reacts with the luminescence solution to emit light, so that detection signals generated by luminescence exist in the sample processing solution quality control site, the sample solution quality control site, the test reaction solution quality control site and the reaction solution quality control site, and the detection signals appear as shown in fig. 5 b.
The technical features of the above embodiments may be arbitrarily combined, and all possible combinations of the technical features in the above embodiments are not described for brevity of description, however, as long as there is no contradiction between the combinations of the technical features, they should be considered as the scope of the description.
The foregoing examples represent only a few embodiments of the present application, which are described in more detail and are not to be construed as limiting the scope of the invention. It should be noted that it would be apparent to those skilled in the art that various modifications and improvements could be made without departing from the spirit of the present application, which would be within the scope of the present application. Accordingly, the scope of protection of the present application is to be determined by the claims appended hereto.
Claims (10)
1. The biochip is characterized by comprising a solid phase carrier, wherein a liquid level quality control microarray is arranged on the solid phase carrier, and comprises at least one of a sample treatment liquid quality control site, a sample liquid quality control site, a test reaction liquid quality control site and a luminescence liquid quality control site; the sample treatment fluid comprises a ruthenium-containing substrate, and a first quality control antibody for specifically binding with the ruthenium-containing substrate is arranged at a quality control site of the sample treatment fluid; the sample liquid quality control site is provided with a second quality control antibody which is used for specifically combining with a non-target object to be detected in the sample liquid; the test reaction solution comprises a detection antibody which specifically binds to a target object to be tested, and a third quality control antibody which is used for specifically binding to the detection antibody is arranged at a quality control site of the test reaction solution; the luminous solution comprises a luminous substrate for carrying out luminous reaction with a first quality control antibody, a second quality control antibody and a detection antibody, at least one of a marker for carrying out luminous reaction with the luminous substrate and a fourth quality control antibody is arranged at a quality control site of the luminous solution, the fourth quality control antibody is not combined with a specific site of the detection antibody on a target object to be detected, the detection antibody and the fourth quality control antibody are both marked with the marker, and the marker is used for carrying out luminous reaction with the luminous substrate.
2. The biochip of claim 1, wherein the liquid level quality control microarray quality control site satisfies at least one of the following features (1) - (6):
(1) The luminous liquid comprises luminol and H 2 O 2 ；
(2) The marker is HRP;
(3) The sample liquid is selected from at least one of plasma, serum, saliva and urine;
(4) The first quality control antibody arranged at the quality control site of the sample treatment fluid is an anti-ruthenium antibody, and the anti-ruthenium antibody is used for specifically combining with a ruthenium-containing substrate in the sample treatment fluid;
(5) The second quality control antibody arranged at the quality control site of the sample liquid is an anti-human IgG antibody.
(6) And the third quality control antibody arranged at the quality control site of the test reaction liquid is an anti-marker antibody.
3. The biochip according to claim 1, wherein the solid support satisfies at least one of the following features (1) to (3):
(1) The solid phase carrier is selected from at least one of a black glass slide, a polystyrene plastic carrier, an NC membrane carrier and a PDVF membrane carrier;
(2) The solid phase carrier is also provided with at least one of a positive quality control microarray, a negative quality control microarray and a test microarray for detecting a target object to be detected.
4. A kit comprising the biochip of any of claims 1-3.
5. The kit of claim 4, further comprising at least one of a sample processing fluid, a sample quality control reaction fluid, a test reaction fluid, and a luminescence fluid.
6. The kit of claim 5, wherein the kit satisfies at least one of the following features (1) to (4):
(1) The sample processing fluid comprises a ruthenium-containing substrate;
(2) The sample quality control reaction liquid comprises an anti-human IgG antibody, and the anti-human IgG antibody in the sample quality control reaction liquid is marked with a marker;
(3) The test reaction liquid comprises a detection antibody and an anti-ruthenium antibody, and the anti-ruthenium antibody in the test reaction liquid is marked with a marker;
(4) The luminous liquid comprises luminol and H 2 O 2 。
7. The kit of claim 6, wherein the kit satisfies at least one of the following features (1) - (2):
(1) The content of the ruthenium-containing substrate in the sample treatment liquid is 5-20 mug/mL;
(2) The anti-human IgG antibodies include at least one of goat anti-human IgG antibodies and mouse anti-human IgG antibodies.
8. A method of detecting a target analyte, the method comprising:
detecting a target analyte in a sample fluid using the kit according to any one of claims 4 to 7;
determining whether the addition amount of at least one of the sample processing liquid, the sample quality control reaction liquid, the test reaction liquid and the luminescent liquid is abnormal in the detection process according to the detection signals of the liquid level quality control microarray of the biochip in the kit.
9. The method according to claim 8, wherein detecting the target analyte in the sample fluid using the kit specifically comprises:
mixing a sample liquid with a sample processing liquid in a kit to obtain a sample mixed liquid, reacting the sample mixed liquid with a biochip in the kit, and reacting the reacted biochip with a test reaction liquid and a luminescent liquid;
and/or
Mixing the sample liquid and the sample processing liquid to obtain a sample mixed liquid, reacting the sample mixed liquid with the biochip in the kit, and reacting the reacted biochip with the sample quality control reaction liquid and the luminescent liquid in the kit.
10. The method according to claim 8 or 9, wherein the sample fluid is selected from at least one of plasma, serum and saliva; and/or
The target object is selected from any one of tumor markers, autoimmune disease markers and cardiopulmonary disease markers.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211732991.3A CN116047059A (en) | 2022-12-30 | 2022-12-30 | Biochip, kit and method for detecting target object to be detected |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211732991.3A CN116047059A (en) | 2022-12-30 | 2022-12-30 | Biochip, kit and method for detecting target object to be detected |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116047059A true CN116047059A (en) | 2023-05-02 |
Family
ID=86112716
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211732991.3A Pending CN116047059A (en) | 2022-12-30 | 2022-12-30 | Biochip, kit and method for detecting target object to be detected |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116047059A (en) |
-
2022
- 2022-12-30 CN CN202211732991.3A patent/CN116047059A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US6921637B2 (en) | Colloid compositions for solid phase biomolecular analytical, preparative and identification systems | |
| EP1348961A1 (en) | Protein chips, method producing it and detection system of the protein chips, and operating method of the detection system | |
| KR20000071894A (en) | Multipurpose diagnostic systems using protein chips | |
| JP4834075B2 (en) | Antigen detection | |
| CN1847850A (en) | Diagnostic control system | |
| JP2002501170A (en) | Apparatus and method for separating at least one specific component in a product sample and apparatus for performing the method | |
| RU2298796C2 (en) | Diagnostic system containing protein chips having features for concurrently determining a plurality of indices | |
| CN116047059A (en) | Biochip, kit and method for detecting target object to be detected | |
| JP6966197B2 (en) | Red blood cell detection | |
| CN116223808A (en) | Biochip, kit and biochip detection method for identifying interference type | |
| US20080230605A1 (en) | Process and apparatus for maintaining data integrity | |
| EP3102953B1 (en) | Crossmatching blood samples | |
| JP2001272406A (en) | Container for high-sensitivity analysis | |
| EP1462801A2 (en) | Methods for determining the negative control value for multi-analyte assays | |
| CN117388499A (en) | Chemiluminescence protein chip method and kit for antigen proteins in serum | |
| US20030224459A1 (en) | Protein detection method | |
| CN113945713A (en) | Biochip for joint detection of multiple tumor markers and preparation and application thereof | |
| US20070037213A1 (en) | Method of analyzing protein and kit for analyzing protein | |
| CN115524485A (en) | Protein chip kit for quantitative quality control of sample adding accuracy and application | |
| CN1261669A (en) | Albumen and gene detecting plate and its preparing method and use | |
| WO2017100454A1 (en) | Automated silver enhancement system | |
| JPWO2017130829A1 (en) | Microarray, microarray manufacturing method, inspection method, and inspection kit | |
| CN106771227A (en) | The method of biological sample bank Rapid identification autoimmune urticaria sample |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |