CN208459404U - Immunochromatography joint detection card - Google Patents

Immunochromatography joint detection card Download PDF

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Publication number
CN208459404U
CN208459404U CN201821120899.0U CN201821120899U CN208459404U CN 208459404 U CN208459404 U CN 208459404U CN 201821120899 U CN201821120899 U CN 201821120899U CN 208459404 U CN208459404 U CN 208459404U
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China
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antibody
immunochromatography
marker
test paper
detection card
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Expired - Fee Related
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CN201821120899.0U
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Chinese (zh)
Inventor
程晓宇
朱世伟
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Accuracy Tianjin Biotechnology Co ltd
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Accuracy Tianjin Biotechnology Co ltd
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Abstract

An immunochromatography joint detection card. The test paper comprises a plastic shell, wherein a first test paper groove and a second test paper groove are formed in the bottom of the plastic shell, a first test paper strip is placed in the first test paper groove, a second test paper strip is placed in the second test paper groove, a first sample inlet hole, a second sample inlet hole, a first detection window and a second detection window are formed in the top of the plastic shell, the first sample inlet hole corresponds to the first test paper groove, the second sample inlet hole corresponds to the second test paper groove, and procalcitonin, interleukin 6, serum amyloid A and C-reactive protein are detected along the first sample inlet hole and the second sample inlet hole. The utility model is used for immunochromatography joint detection.

Description

A kind of immunochromatography combined detection card
Technical field:
The utility model relates to a kind of immunochromatography combined detection cards.
Background technique:
Lateral flow detection method is also referred to as lateral flow immunochromatographic measurement, is for target analytes in test sample The portable unit that whether there is, can use, can be equipped with small and exquisite in the case where not needing special and expensive equipment Portable detection device is as support;Under normal conditions, these reagent facilities are used for the medicine of home test, emergency treatment or laboratory Diagnosis;One wide-scale distribution and well-known application example the most is home-use pregnancy test card.
Sample pad in immunochromatography plays the role of sponge, contains excessive sample liquids;Once being saturated, liquid Flowing transport is into labeling pad, the marker that store in labeling pad, is dry biologically active granulated Formula, keeps activity in the matrix such as salt and sugar, and with detected target molecule specificity can occur for these active materials Reaction, as the compound of the immune combination of antibody antigen, these reactions has been fixed at the surface of the particles.When sample liquids dissolve When salt saccharide matrix, it can also dissolve in particle wherein, and while flowing through porous structure film, and be fixed in the above in advance Other ligand binding, the band being formed on fixed position, there are two types of band is general, test strip shows detected target The content situation of object, whether Quality Control band represents flowing reactive state good.Final liquid, which flows into, to be absorbed in phase, test It completes.
C reactive protein is typical acute phase protein in inflammatory reaction;It is synthesized by liver and by five identical polypeptide chain Composition forms the five-membered ring that molecular weight is 105000 dalton;CRP be it is most sensitive in acute phase reactant, in inflammation mistake Its concentration increases sharply in journey;Compound CRP activates the complement system started with C1q;Then the conditioning of CRP starting invasion cell is made With and phagocytosis, but its major function be combine and detoxify by tissue damage generate endogenous noxious material;CRP detection is used In detection systemic inflammatory process;Assessment antibiotic treatment bacterium infection;It is broken with early stage amnion simultaneously to detect intrauterine infection It splits;Distinguish the disease of the active and inactive form of infection simultaneously, for example, in the patient for suffering from SLE or ulcerative colitis into Row;It therapeutically monitors rheumatic disease and assesses anti-inflammatory therapy;It determines that there are postoperative complications in early stage, such as infects Wound, thrombosis and pneumonia, and distinguish infection and marrow graft rejection reaction;High quick CRP test has been used for early detection Pediatrics and coronary heart disease risk assessment.Some researchs are concluded that highly sensitive CRP detection may be used as pre- detect The index of aobvious healthy population coronary heart disease risk, and the index as prediction recurrent events;The increase of CRP value be it is nonspecific, It should not be explained in the case where no complete clinical medical history.
Interleukin-6 (IL-6) is the pleiotropic cytokines with wide variety of functions;It is described as interfering first Element-β 2, plasmacytoma growth factor and hepatocyte-stimulating factor;It is described as human B cell stimulating factor 2(BSF2 later).? 1988, it is thus proposed that IL-6 is named as, because further the verified protein of research not only shows B cell Activity, and show T cell, candidate stem cell, the activity of liver cell and brain cell;One coding, 212 amino acid products Term single gene is generated the peptide of 184 amino acid of the molecular weight between 22-27 kDa in the end N- by cutting.1989, It is reported that the immunocompetence compound there are also some 60-70kDa is detected in the body fluid of acute bacterial infection patient;With Damage, wound, pressure, infection, brain death, tumor are formed during acute inflammatory reaction relevant with other situations, and IL-6 is fast Speed induction generates;The IL-6 concentration of detection trauma patient can predict the late complication of more Fundamental Operations, or instruction It fails to pinpoint a disease in diagnosis or complication;The continuous measurement of IL-6 in the serum or blood plasma of the patient of ICU is moved in, display can be used for assessing SIRS(complete Body inflammatory response syndrome), the severity of pyemia and infectious shock, and predict the prognosis of these patients;IL-6 can also As Early warning indicator for detecting sepsis of the newborn;IL-6 also works in chronic inflammation, such as rheumatoid arthrosis It is scorching.
The prohormone that Procalcitonin (PCT) is made of 116 amino acid, molecular weight are about 12.7 kD;PCT is by mind Through endocrine cell (the C cell including thyroid gland, lung and pancreatic tissue) express, through digestion be decomposed into (prematurity) calcitonin, C-terminal peptides and amino terminal peptide.Only contain a small amount of PCT in Healthy People blood;It has been found that PCT can be significantly raised after bacterium infection.It is dynamic When object model test shows that septicopyemia occurs for body, Various Tissues may express PCT;The intracorporal PCT of patients with sepsis 114 amino acid are contained only, the Ala-Pro of amino terminal is lacked;The raising of PCT level sees bacillary septicopyemia, especially Severe septicopyemia and infectious shock;PCT can be used as the prognostic indicator of patients with sepsis;It is also acute critical pancreatitis And its reliability index of major complications;For Community Acquired Respiratory Tract Infection and air-conditioning inductivity patients with pneumonia, PCT can make For the index of antibiotic selection and curative effect judgement.
Serum amyloid A protein (SAA) protein includes (12-14kDa, the 104-112 amino acid of a small-molecular-weight Residue) family is the protein of highly conserved differential expression in vertebrate;SAA albumen participates in acute phase response;This A little reactions are the direct host responses for early stage being directed to inflammation;In acute stage, recycles SAA level and increase 100-1000 times, concentration Reach 1 milligram every milliliter;SAA further relates to several morbid states, including rheumatoid arthritis, atherosclerosis, AA starch Sample denaturation and coronary artery disease;Liver is the main portions of SAA synthesis, expresses outside liver and also has been reported that;Mankind's SAA gene family It is positioned at No. 11 chromosomes;In human body, identified four kinds of SAA genes and three kinds of protein products: mankind SAA1 and SAA2 is designated as acute stage SAA(A-SAA) isotype;And SAA4 is composition expression type, and SAA3 is pseudogene;A-SAA Belong to a kind of acute phase protein, further includes C reactive protein (CRP), haptoglobin, ceruloplasmin, complement in this type Ingredient C3 and C4, α 1- acidoglycoprotein, Prolastin and fibrinogen.In human body, CRP and SAA be all Acute stage synthesis;And in mouse, SAA is main acute phase protein.
As the albumen of inflammatory reaction acute stage, C reactive protein, Procalcitonin, interleukin-6, serum amyloid sample Albumin A has important auxiliary diagnosis meaning early stage infection and in the development process of inflammation;Meanwhile in Other diseases state In, also marker also will appear measurable apparent increase for this;Therefore, the joint-detection of these markers is very significant 's;There is presently no reagents can detect these four markers simultaneously, clinically needs to separate progress, or even need using different Detection method reagent carry out, if Procalcitonin, interleukin-6 use immunochromatography or chemiluminescent method, together When C reactive protein, serum amyloid A protein detected using the method for immunoturbidimetry.Even same detection method, The detection of these four markers also largely needs separately to carry out, or needs the cooperation of the fully-automatic equipment of higher cost.
Summary of the invention:
The purpose of the utility model is to provide a kind of using easy to operate, detection process is quick, joint-detection is high-efficient A kind of immunochromatography combined detection card.
Above-mentioned purpose is realized by following technical scheme:
A kind of immunochromatography combined detection card, composition includes: plastic shell, and the bottom of the plastic shell is provided with No.1 test paper slot and No. two test paper slots, the No.1 test paper slot is interior to place test strips one, places in No. two test paper slots Test strips two are provided with No.1 sample holes, No. two sample holes, No.1 detection window, No. two detections at the top of the plastic shell Window, the corresponding No.1 test paper slot of the No.1 sample holes, corresponding No. two test paper of No. two sample holes Slot detects Procalcitonin, interleukin 6, serum amyloid A protein along the No.1 sample holes and No. two sample holes And C reactive protein.
A kind of immunochromatography combined detection card, the test strips one include No.1 PVC board, the No.1 No.1 nitrocellulose filter is placed in PVC board, placement No.1 labeling pad on the No.1 nitrocellulose filter, described one No.1 sample pad is placed on labelled notation object pad, places No.1 blotting paper on the No.1 nitrocellulose filter.
A kind of described immunochromatography combined detection card, the test strips two include No. two PVC boards, and described No. two No. two nitrocellulose filters are placed in PVC board, place two labelled notation object pads on No. two nitrocellulose filters, described two No. two sample pads are placed on labelled notation object pad, place No. two blotting papers on No. two nitrocellulose filters.
A kind of described immunochromatography combined detection card, the marker in the No.1 labeling pad is Procalcitonin Antibody marker and interleukin 6 antibody marker are coated with Procalcitonin antibody on the No.1 nitrocellulose filter respectively T1 detection line, interleukin 6 antibody T2 detection line, two antiantibody C1 nature controlling lines.
A kind of described immunochromatography combined detection card, the marker on the two labelled notation object pads is serum amyloid Sample protein A antibody marker and C reactive protein antibody marker are coated with blood on No. two nitrocellulose filters respectively Clear amyloid A antibody T3 detection line, C reactive protein antibody T4 detection line, two antiantibody C2 nature controlling lines.
The material of a kind of immunochromatography combined detection card, No.1 labeling pad and two labelled notation object pads is glass One of cellulose membrane, polyester or non-woven fabrics, the tracer marked on antibody marker are that colloidal gold, fluorescein, fluorescence are micro- One of ball, color micro-sphere or combination, colloidal gold, fluorescein, fluorescent microsphere, color micro-sphere content be respectively 0.00005mg-0.05mg。
A kind of immunochromatography combined detection card, two antiantibody C1 nature controlling lines and two antiantibody C2 nature controlling lines are needle To colloidal gold, fluorescein, fluorescent microsphere, color micro-sphere antibody secondary antibody, for anti-mouse, anti-sheep, anti-rabbit, anti-chicken secondary antibody one kind Or combination.
A kind of immunochromatography combined detection card, the No.1 nitrocellulose filter and No. two nitric acid The content of coated four kinds of antibody and secondary antibody is respectively 0.0001mg-0.01mg on cellulose membrane.
The width of a kind of immunochromatography combined detection card, test strips one and test strips two is 0.2cm-0.8cm, Length is 5.5cm-10cm.
The utility model has the advantages that
1. the utility model being incorporated into the multiple projects for being used to infect detection in a kind of detection reagent, improve The efficiency of detection, reduces waste, convenient, flexible.
The utility model reduces the erroneous judgement that a kind of reagent result increases generation as caused by non-infective agent, the side of passing through Just quick joint-detection improves the judgement efficiency of clinical application.Manpower, equipment and time cost are reduced to a certain extent.
Detailed description of the invention:
Attached drawing 1 is the structural schematic diagram of this product.
Attached drawing 2 is the top view of this product detection card.
Specific embodiment:
Below in conjunction with the attached drawing of the utility model, the technical scheme in the embodiment of the utility model is carried out clear, complete Site preparation description.
Embodiment 1:
A kind of immunochromatography combined detection card, composition includes: plastic shell 1, and the bottom of the plastic shell is opened There are a No.1 test paper slot 2 and No. two test paper slots 3, place test strips 1 in the No.1 test paper slot, in No. two test paper slots Place test strips 25, be provided at the top of the plastic shell 6, No. two sample holes 7 of No.1 sample holes, No.1 detection window 24, No. two detection windows 25, the corresponding No.1 test paper slot of the No.1 sample holes, described in No. two sample holes are corresponding No. two test paper slots, detect Procalcitonin, interleukin 6, serum along the No.1 sample holes and No. two sample holes Amyloid A and C reactive protein.
Embodiment 2:
The immunochromatography combined detection card of one kind described in embodiment 1, the test strips one include No.1 PVC board 8, institute No.1 nitrocellulose filter 9 is placed in the No.1 PVC board stated, and places No.1 labeling pad on the No.1 nitrocellulose filter 10, No.1 sample pad 11 is placed in the No.1 labeling pad, and No.1 water suction is placed on the No.1 nitrocellulose filter Paper 12.
Embodiment 3:
The immunochromatography combined detection card of one kind described in embodiment 1, the test strips two include No. two PVC boards 13, No. two nitrocellulose filters 14 are placed in No. two PVC boards, place two labelled notations on No. two nitrocellulose filters Object pad 15 places No. two sample pads 16 on the two labelled notation object pads, places No. two on No. two nitrocellulose filters Blotting paper 17.
Embodiment 4:
The immunochromatography combined detection card of one kind as described in example 2, the marker in the No.1 labeling pad are Procalcitonin antibody marker and interleukin 6 antibody marker are coated with drop calcium on the No.1 nitrocellulose filter respectively Plain original antibody T1 detection line 18, interleukin 6 antibody T2 detection line 19, two antiantibody C1 nature controlling lines 20.
Embodiment 5:
The immunochromatography combined detection card of one kind described in embodiment 3, the marker on the two labelled notation object pads are Serum amyloid A protein antibody marker and C reactive protein antibody marker, on No. two nitrocellulose filters respectively It is coated with serum amyloid A protein antibody T3 detection line 21, C reactive protein antibody T4 detection line 22, two antiantibody C2 nature controlling lines 23。
Embodiment 6:
The immunochromatography combined detection card of one kind described in embodiment 2 or 3, No.1 labeling pad and two labelled notation object pads Material is one of glass fibre element film, polyester or non-woven fabrics, and the tracer marked on antibody marker is colloidal gold, fluorescence One of element, fluorescent microsphere, color micro-sphere or combination, the content difference of colloidal gold, fluorescein, fluorescent microsphere, color micro-sphere For 0.00005mg-0.05mg.
Embodiment 7:
The immunochromatography combined detection card of one kind described in embodiment 4 or 5, two antiantibody C1 nature controlling lines and two antiantibody C2 Nature controlling line be for colloidal gold, fluorescein, fluorescent microsphere, color micro-sphere antibody secondary antibody, for anti-mouse, anti-sheep, anti-rabbit, anti-chicken One kind or combination of secondary antibody.
Embodiment 8:
The immunochromatography combined detection card of one kind described in embodiment 2 or 3, No.1 nitrocellulose filter and No. two nitric acid fibres Tieing up the content of coated four kinds of antibody and secondary antibody on plain film is respectively 0.0001mg-0.01mg.
Embodiment 9:
The width of the immunochromatography combined detection card of one kind described in embodiment 1, test strips one and test strips two is 0.2cm-0.8cm, length 5.5cm-10cm.
Embodiment 10:
The immunochromatography combined detection card of one kind described in above-described embodiment, by test strips one, test strips two and plastics outside Shell composition, test strips are made of No.1 PVC board and No. two PVC boards, sample pad, labeling pad, nitrocellulose filter, blotting papers. In test strips one, marker is Procalcitonin antibody marker and interleukin 6 antibody marker, is wrapped respectively on nitrocellulose filter There are Procalcitonin antibody (T1 detection line), interleukin 6 antibody (T2 detection line), two antiantibodys (C1 nature controlling line).Test strips two In, marker is serum amyloid A protein antibody marker and C reactive protein antibody marker, is distinguished on nitrocellulose filter It is coated with serum amyloid A protein antibody (T3 detection line), C reactive protein antibody (T4 detection line), (the C2 Quality Control of two antiantibodys Line).Test strips one and test strips two are put into two test paper slots of plastic shell, there is two sample holes on plastic shell respectively Sample-adding, while detecting Procalcitonin, interleukin 6, serum amyloid A protein and C reactive protein.This combined detection card have make With the feature that easy to operate, detection process is quick, joint-detection is high-efficient.
Embodiment 11:
The immunochromatography combined detection card of one kind described in above-described embodiment, the preparation of immunochromatography combined detection card;Make With four kinds of label antibody of colloid gold label, it is dissolved in buffer, specifically: sucrose 2g/L, BSA 0.5g/L, 20mM phosphorus Phthalate buffer solution, pH=7.4;Concentration of ordinary dissolution is 2ug/ml;Procalcitonin, interleukin 6, which are uniformly mixed, is coated on glass On cellulosic mat;Serum amyloid A protein and C reactive protein, which are uniformly mixed, to be coated on another glass fibre element pad;It dried It is cut after night.
It is sprayed respectively on position in the detection line T1 and detection line T2 of the nitrocellulose filter in No.1 PVC board, nature controlling line C1 Apply Procalcitonin antibody, interleukin 6 antibody and secondary antibody;The detection line T3 and T4 of nitrocellulose filter in No. two PVC boards, matter Control sprays serum amyloid A protein antibody, C reactive protein antibody and secondary antibody on the position line C2 respectively;The concentration of spray solution is 1mg/ml, quantity for spray 1ul/cm.
After sample pad and water suction paper wood cutting, it is pasted onto together with labeling pad in No.1 PVC board and No. two PVC boards.
The No.1 PVC board pasted and No. two PVC boards are cut into test strips, and are put into plastic shell test paper slot, is pressed Tight plastic shell.
The detection card installed is encapsulated together with desiccant, storage is stand-by.
Embodiment 12:
The immunochromatography combined detection card of one kind described in above-described embodiment, by test strips one, test strips two and plastics outside Shell composition, test strips are made of PVC board, sample pad, labeling pad, nitrocellulose filter, blotting paper.
The described plastic shell tool is there are two the slot for placing test paper and two corresponding sample holes, test strips one and examination Paper slip two is put into two test paper slots, is loaded respectively by two sample holes on plastic shell, while detecting the drop calcium in sample Plain original, interleukin 6, serum amyloid A protein and C reactive protein.The width of test strips is 0.2cm-0.8cm, and length is 5.5cm-10cm。
Preferably, the mat material matter of labeling pad is one of glass fibre element film, polyester or non-woven fabrics.
Preferably, the tracer marked on antibody marker is colloidal gold, in fluorescein, fluorescent microsphere, color micro-sphere A kind of or combination, it is further preferred that tracer is one of fluorescein, fluorescent microsphere or combination.
In the test strips one, marker is Procalcitonin antibody marker and interleukin 6 antibody marker, and nitric acid is fine It ties up and is coated with Procalcitonin antibody (T1 detection line), interleukin 6 antibody (T2 detection line), two antiantibodys (C1 matter respectively on plain film Control line), it is preferable that the content of antibody marker is respectively 0.00005mg-0.05mg, and two antiantibodys are for four kinds of markers The secondary antibody of antibody, for anti-mouse, anti-sheep, anti-rabbit, anti-chicken secondary antibody one kind or combination, on nitrocellulose filter coated four kinds it is anti- The content of body and secondary antibody is respectively 0.0001mg-0.01mg.
In the test strips two, marker is serum amyloid A protein antibody marker and C reactive protein antibody mark Remember object, is coated with serum amyloid A protein antibody (T3 detection line), C reactive protein antibody (T4 respectively on nitrocellulose filter Detection line), two antiantibodys (C2 nature controlling line), it is preferable that the content of antibody marker is respectively 0.00005mg-0.05mg, secondary antibody Antibody is the secondary antibody for four kinds of marker antibodies, for anti-mouse, anti-sheep, anti-rabbit, anti-chicken secondary antibody one kind or combination, nitric acid is fine Tieing up the content of coated four kinds of antibody and secondary antibody on plain film is respectively 0.0001mg-0.01mg.
This product it is practical to all reagents, material be commercial goods, be easier to obtain, such as antibody C reactive protein and Interleukin-6, secondary antibody can be obtained in Meridian Life Science company, such as antibody Procalcitonin and serum amyloid sample Albumin A can be obtained in Hytest company, if fluorescein and fluorescent microsphere can be obtained in LIFE company, as other materials all can be MILLIPORE company obtains, and such as other reagents can all be obtained in Sigma company, or obtains respectively in different company.
Immunochromatography combined detection card described in this product can be made by following steps:
The antibody for being used to mark is coated in marker mat material, is cut after dry in conjunction with tracer-labelling respectively.
Nitrocellulose filter is pasted onto PVC board (No.1 PVC board and No. two PVC boards), coated antibody point will be used for It is not coated on the corresponding position of nitrocellulose filter, and dry.
After sample pad and water suction paper wood cutting, it is pasted onto PVC board together with labeling pad.
The PVC board pasted is cut into test strips, and is put into plastic shell test paper slot, plastic shell is compressed.
The detection card installed is encapsulated together with desiccant, storage is stand-by.

Claims (9)

1. a kind of immunochromatography combined detection card, composition includes: plastic shell, it is characterized in that: the plastic shell Bottom is provided with No.1 test paper slot and No. two test paper slots, places test strips one, No. two test paper in the No.1 test paper slot Test strips two are placed in slot, be provided at the top of the plastic shell No.1 sample holes, No. two sample holes, No.1 detection window, No. two detection windows, the corresponding No.1 test paper slot of the No.1 sample holes, No. two sample holes correspond to described No. two test paper slots form sediment along the No.1 sample holes and No. two sample holes detection Procalcitonin, interleukin 6, serum Powder sample albumin A and C reactive protein.
2. the immunochromatography combined detection card of one kind according to claim 1, it is characterized in that: the test strips one are wrapped No.1 PVC board is included, No.1 nitrocellulose filter is placed in the No.1 PVC board, is put on the No.1 nitrocellulose filter No.1 labeling pad is set, No.1 sample pad is placed in the No.1 labeling pad, is put on the No.1 nitrocellulose filter Set No.1 blotting paper.
3. the immunochromatography combined detection card of one kind according to claim 1, it is characterized in that: the test strips two are wrapped No. two PVC boards are included, No. two nitrocellulose filters are placed in No. two PVC boards, are put on No. two nitrocellulose filters Two labelled notation object pads are set, No. two sample pads are placed on the two labelled notation object pads, are put on No. two nitrocellulose filters Set No. two blotting papers.
4. the immunochromatography combined detection card of one kind according to claim 2, it is characterized in that: the No.1 marker Marker on pad is Procalcitonin antibody marker and interleukin 6 antibody marker, on the No.1 nitrocellulose filter It is coated with Procalcitonin antibody T1 detection line, interleukin 6 antibody T2 detection line, two antiantibody C1 nature controlling lines respectively.
5. the immunochromatography combined detection card of one kind according to claim 3, it is characterized in that: the two labelled notation object pads On marker be serum amyloid A protein antibody marker and C reactive protein antibody marker, No. two nitric acid fibres It ties up and is coated with serum amyloid A protein antibody T3 detection line, C reactive protein antibody T4 detection line, two antiantibodys respectively on plain film C2 nature controlling line.
6. the immunochromatography combined detection card of one kind according to claim 2 or 3, it is characterized in that: No.1 labeling pad and The material of two labelled notation object pads is one of glass fibre element film, polyester or non-woven fabrics, the tracer marked on antibody marker Object is one of colloidal gold, fluorescein, fluorescent microsphere, color micro-sphere or combination, colloidal gold, fluorescein, fluorescent microsphere, colour The content of microballoon is respectively 0.00005mg-0.05mg.
7. the immunochromatography combined detection card of one kind according to claim 4 or 5, it is characterized in that: two antiantibody C1 Quality Controls Line and two antiantibody C2 nature controlling lines be for colloidal gold, fluorescein, fluorescent microsphere, color micro-sphere antibody secondary antibody, for anti-mouse, anti- Sheep, anti-rabbit, anti-chicken secondary antibody one kind or combination.
8. the immunochromatography combined detection card of one kind according to claim 2 or 3, it is characterized in that: No.1 nitrocellulose The content of coated four kinds of antibody and secondary antibody is respectively 0.0001mg-0.01mg on film and No. two nitrocellulose filters.
9. the immunochromatography combined detection card of one kind according to claim 1, it is characterized in that: test strips one and test strips Two width is 0.2cm-0.8cm, length 5.5cm-10cm.
CN201821120899.0U 2018-07-16 2018-07-16 Immunochromatography joint detection card Expired - Fee Related CN208459404U (en)

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