CN111103280A - Test strip for detecting insulin-resistant autoantibody and preparation method thereof - Google Patents
Test strip for detecting insulin-resistant autoantibody and preparation method thereof Download PDFInfo
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- CN111103280A CN111103280A CN201911362124.3A CN201911362124A CN111103280A CN 111103280 A CN111103280 A CN 111103280A CN 201911362124 A CN201911362124 A CN 201911362124A CN 111103280 A CN111103280 A CN 111103280A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
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- 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
- G01N33/52—Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements
Abstract
The invention relates to the field of immunodetection, and discloses a test strip for detecting an insulin-resistant autoantibody, which comprises a PVC (polyvinyl chloride) base plate, and a sample pad, a combined release pad, a nitrocellulose membrane and an absorption pad which are sequentially lapped on the PVC base plate, wherein the combined release pad contains anti-human IgG marked by colloidal gold, and the nitrocellulose membrane is provided with a quality control line coated by goat anti-mouse IgG antibody and a detection line coated by insulin antigen. The invention also provides a preparation method of the test strip. The test strip has strong specificity and does not have cross reaction with other autoantibodies; the sensitivity detection result shows stable property and good reproducibility. The method is simple, convenient to operate and has good application prospect.
Description
Technical Field
The invention relates to the field of immunodetection, in particular to a test strip for detecting an anti-insulin autoantibody and a preparation method thereof.
Background
Diabetes is a common disease caused by the interaction of genetic and environmental factors, hyperglycemia is taken as a main clinical sign, common symptoms comprise polydipsia, diuresis, polyphagia, emaciation and the like, and the diabetes is divided into type 1 diabetes and type 2 diabetes.
Human Leukocyte Antigen (HLA) genes are located on the 6 th chromosome short arm, and form a group of closely linked genes, and HLA is coded by I, II and III 3 genes. The class I gene region comprises HLA-A, HLA-B, HLA-C and other genes with unknown functions and pseudogenes, and antigen molecules coded by the gene are present on the surfaces of all nucleated cells and are responsible for presenting foreign antigens to T lymphocytes of CD 8; the II-class gene region mainly comprises 3 subregions of HLA-DR, HLA-DQ and HLA-DP, respectively encodes DR, DQ and DP antigens, is present on the surfaces of mature B lymphocytes and antigen presenting cells and is responsible for presenting the antigens to CD4 cells; the class III gene region encodes some soluble proteins including some complement components, such as C2C4A, C4B, Tumor Necrosis Factor (TNF), and Heat Shock Protein (HSP). HLA is restricted by Major Histocompatibility Complex (MHC), is involved in the interaction of T lymphocytes in recognizing antigens and other immune cells, and in the formation and maintenance of self tolerance, and plays an important role in recognizing self and heterosis, inducing and regulating immune responses, and the like. As can be seen, HLA is associated with the development of many autoimmune diseases, including type 1 diabetes.
It is well established that type 1 diabetes is caused by immune-mediated islet β cell destruction, and it has been demonstrated that during the development of type 1 diabetes, a variety of autoantibodies (IAAs) against islet β cells can be detected, the positive rate of insulin autoantibodies IAA is about 50% in diagnosed type 1 diabetes patients.
Disclosure of Invention
The invention aims to provide a test strip for detecting an anti-insulin autoantibody, which comprises a PVC (polyvinyl chloride) base plate, a sample pad, a combination release pad, a nitrocellulose membrane and an absorption pad, wherein the sample pad, the combination release pad, the nitrocellulose membrane and the absorption pad are sequentially lapped on the PVC base plate, the combination release pad contains anti-human IgG marked by colloidal gold, and the nitrocellulose membrane is provided with a quality control line coated by a second anti-human IgG antibody and a detection line coated by insulin antigen.
Preferably, the anti-human IgG is of murine origin. More preferably, the anti-human IgG secondary antibody is a goat anti-mouse secondary antibody.
The invention applies the antigen-antibody reaction and the immunochromatography one-step principle, namely, an anti-insulin autoantibody (IAA) in serum is combined with anti-Human IgG marked by colloidal gold in a detection reagent and then is combined with insulin antigen fixed on a nitrocellulose membrane, so that a detection color band is formed by deposition, and a colorless band is displayed at a Control-line (C-line) which shows the Test paper effective according to a Test-line (T-line for short) in the middle section of the Test paper and above the Test-line, and the Test paper is used as rapid detection Test paper for the auxiliary diagnosis of insulin-dependent diabetes (type 1 diabetes).
The invention also provides a kit for detecting the anti-insulin autoantibody, which comprises the test strip.
The invention also provides a preparation method of the test strip, which comprises the following steps:
(1) preparing a colloidal gold solution;
(2) carrying out colloidal gold labeling on the anti-human IgG antibody; mixing the colloidal gold and the anti-human IgG antibody, centrifuging, and carrying out heavy suspension precipitation by using a PBS (phosphate buffer solution) solution to obtain an anti-human IgG-colloidal gold compound;
(3) preparing a gold-labeled combined release pad: uniformly spraying an anti-human IgG-colloidal gold compound on the combined release pad;
(4) preparing a nitrocellulose membrane: respectively coating the anti-human IgG secondary antibody and the insulin antigen on a quality control line and a detection line on a nitrocellulose membrane;
(5) assembling the test strip: and adhering the sample pad, the combined release pad, the coated nitrocellulose membrane and the absorption pad on a PVC plate in a dry environment, and cutting to obtain the test strip.
Preferably, the anti-human IgG is of murine origin. More preferably, the anti-human IgG secondary antibody is a goat anti-mouse secondary antibody.
The test strip for the insulin-resistant autoantibody is simple in detection method and simple and convenient to operate; the specificity is strong, and the cross reaction with other autoantibodies is avoided; sensitivity and batch-to-batch precision detection, stable property of result display, good reproducibility and good application prospect.
Description of the drawings:
FIG. 1 is a schematic structural diagram of the test strip of the present invention.
1-sample pad
2-binding release pad containing colloidal gold labeled anti-human IgG
3-nitrocellulose membrane coated with insulin antigen and goat anti-mouse IgG antibody
4-absorbent pad
5-PVC soleplate.
The specific implementation mode is as follows:
the invention discloses an anti-insulin autoantibody detection test strip, which can be realized by appropriately improving process parameters by referring to the content in the text by a person skilled in the art. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the methods and applications of this invention have been described in terms of preferred embodiments, it will be apparent to those of ordinary skill in the art that variations and modifications in the methods and applications described herein, as well as other suitable variations and combinations, may be made to implement and use the techniques of this invention without departing from the spirit and scope of the invention.
Example 1:
1) intermediate solution preparation
1. Preparation of 0.2M Potassium carbonate solution
2. Preparation of 1% sodium citrate solution
3. Preparation of 1% chloroauric acid solution
4. Preparation of 10% bovine serum albumin solution
5. Method for preparing 0.1M tris-HCL solution with pH of 8.0-8.5
6. Method for preparing 0.1M phosphate buffer solution
1. Preparation of colloidal gold solution
1.1 preparation of the device:
electronic temperature-control magnetic stirring electric heating sleeve pipette gun measuring cylinder
1.2 formula:
1ml of 1% chloroauric acid solution was added to 100ml of water, and 1.0-1.8ml of 1% sodium citrate solution was added after boiling.
1.3, operation steps:
1.3.1 measuring and preparing laboratory tertiary water with a certain volume;
1.3.2 completely dissolving 1% chloroauric acid solution in a certain proportion in laboratory tertiary water, uniformly mixing and heating to boil;
1.3.3 adding 1% citric acid solution, stirring to change the color of the solution until the color does not change, stopping stirring and heating, and naturally cooling to room temperature.
2. Mouse anti-human IgG antibody labeling and centrifugation
2.1 preparation of the device:
liquid-transfering gun, flask, electronic temp. -controlled magnetic stirring electric heating jacket and centrifugal machine
2.2 formula:
adding 2-6 microliters of 0.2M potassium carbonate solution into 1 milliliter of colloidal gold solution, shaking and uniformly mixing, and adding 10-15 microliters of 1.0mg/ml potassium carbonate solution
The mouse anti-human IgG antibody is mixed evenly, 10% BSA 100ul is added into 1ml colloidal gold solution of the labeled protein as a stabilizing agent, and the solution is dissolved by one tenth of the volume before redissolution after centrifugation.
2.3, operation steps:
2.3.1 taking colloidal gold solution which is qualified in inspection.
2.3.2 adding a calculated and determined amount of 0.2M potassium carbonate solution under stirring, then adding 1.0mg/ml of mouse anti-human IgG antibody and uniformly mixing; rest for 20 minutes.
2.3.3 adding 10% bovine serum albumin solution according to the calculated amount, uniformly mixing and standing for 20 minutes;
2.3.4 then put into a centrifuge tube, centrifuge at 10000rpm for 10min, and discard the supernatant.
2.3.5 the precipitate was dissolved with a calculated amount of tris-HCL pH8.0-8.5 redissolution.
3. IAA metal spraying
3.1 required appliances:
three-dimensional plane film-drawing instrument reagent bottle liquid-transferring gun
3.2 Process conditions:
drying is carried out in a drying box with the relative humidity of less than or equal to 40 percent at the temperature of 35-40 ℃.
3.3, operation steps:
3.3.1 put the gold spraying pipette of the three-dimensional plane film scribing instrument into the colloidal gold solution of the labeled protein.
3.3.2 spreading the glass fiber on the instrument, setting the spraying amount of 8-12 mul/cm, and spraying gold.
3.3.3 placing the sprayed glass fiber on a test tube rack, placing the test tube rack in a drying box, drying for 5-8 hours at the temperature of 35-40 ℃ and the relative humidity of less than or equal to 40%.
4. IAA coating
4.1 required appliances:
continuous film-drawing instrument reagent bottle liquid-transferring gun
4.2 Process conditions
Drying is carried out in a drying box with the relative humidity of less than or equal to 40 percent at the temperature of 35-40 ℃.
4.3, operation steps:
4.3.1 pipette amounts of insulin antigen and goat anti-mouse secondary antibody calculated according to the production amount were measured in reagent bottles, respectively.
4.3.2 insulin antigen was diluted to 0.8-2.0mg/ml and goat anti-mouse secondary antibody was diluted to 0.8-2.0mg/ml with 0.1M phosphate buffer, respectively.
4.3.3 tear off the end lining paper of PVC plate, align underline and paste the nitrocellulose membrane of the equivalent width, then lay it on the continuous film-drawing instrument and draw line, set the concentration of drawing line to be 0.8-1.5 μ l/cm, the first pipeline is coated with insulin antigen, the second pipeline is coated with goat anti-mouse second antibody.
4.3.4 placing the sprayed PVC board on a test tube rack, placing the test tube rack in a drying oven, drying for 5-8 hours at 35-40 ℃ and with the relative humidity less than or equal to 40%.
5. IAA large panel assembly
5.1 required instruments:
paper cutter
5.2, operation steps:
6.2.11 the large test paper is prepared by sticking adhesive sticker to the hand-held end and sticking adhesive sticker marked with MAX line to the sample-adding end, and cutting into test paper strips with specified width. Before cutting into strips in batch, 10 strips are cut, and 3 strips are randomly spot-checked and measured by production personnel, wherein the width error of a normal test strip is not more than +/-0.04 mm. Card type: and (4) placing the test paper into a clamping and molding part, and pressing the shell by using a shell pressing machine.
6.2.2 bagging
6.2.2.1 each test paper is put into a drying agent and one test paper strip in an aluminum plastic bag. The card type adds the straw one.
6.2.2.2 the aluminum plastic bag is sealed by heat sealing on a multifunctional automatic film sealing machine at the temperature of 180-240 ℃.
Example 2: quality control standard
1) Raw material quality standard
1.1 appearance
The antibody is clear and transparent liquid observed by naked eyes, and does not contain foreign matters or turbid or non-scattering sediment.
1.2 purity and molecular weight:
the molecular weight of the mouse anti-human IgG antibody protein is mainly concentrated on about 25KD, and the purity is more than or equal to 95 percent
The purity of the goat anti-mouse IgG secondary antibody is more than or equal to 90 percent
1.3 protein concentration
The content of insulin antigen is more than or equal to 27U/ml
The mouse anti-human IgG antibody is more than or equal to 1.0mg/ml, and the goat anti-mouse IgG secondary antibody is more than or equal to 1.0 mg/ml.
1.4 titer:
murine anti-human IgG antibody titers 1: 64
Goat anti-mouse IgG secondary antibody 1: 64
1.5 biological Activity
The general method of preparing colloidal gold is to coat the detection line with insulin antigen, coat the quality control line with goat anti-mouse IgG antibody, label the colloidal gold with mouse anti-human IgG antibody, prepare a reagent sample, and meet the requirements of detection sensitivity, specificity and stability.
2) Standard of acceptance of auxiliary materials
1 nitrocellulose membrane
Thickness: 215 +/-20 um.
Capillary migration rate: the chromatography time of water per 4cm was 135. + -.30 s.
2. Glass cellulose membrane
Appearance: the surface is smooth, no stain is generated, and the fiber structure is uniform and compact.
Thickness: and (3) gold pad gluing: 0.5 +/-0.05 mm.
Liquid moving speed: the moving speed is more than or equal to 5mm multiplied by 70mm/min
3. Other auxiliary materials
PVC board, absorbent filter paper, MAX line label, color leather label, etc. are purchased from qualified manufacturers, and corresponding acceptance standard is established.
1. Quality standard of colloidal gold solution
Appearance: rose transparent liquid.
Absorbance: the absorbance of A395/A405 is more than or equal to 1.
2. Marking, centrifugation quality standards
Appearance after gold spraying and drying: smooth and clean, no damage and no pollution. The original rose color of the gold-labeled mouse anti-human IgG antibody is kept, and the color change cannot occur.
And (3) performance detection:
and (3) sensitivity detection: blank control solution: the phosphate buffer solution containing bovine serum albumin was not detected to develop color. Detecting a sample solution containing 50mU/mLIAA standard: only the quality control line is colored, and the detection line is not colored. Detecting a sample solution containing 150mU/mLIAA standard: the quality control line and the detection line are colored. Detecting a sample solution containing 300mU/mLIAA standard: the quality control line and the detection line are colored.
And (3) specific detection: does not cross-react with other autoantibodies.
And (3) detecting the precision in the batch: detecting a sample solution containing 50mU/mLIAA standard: the reaction results are consistent, and the color development is uniform. Detecting a sample solution containing 150mU/mLIAA standard: the reaction results are consistent, and the color development is uniform. Detecting a sample solution containing 300mU/mLIAA standard: the reaction results are consistent, and the color development is uniform.
3. Quality standard of semi-finished product
Appearance: the adhesive is flat and clean, has no damage and pollution, has complete components, correct position, firm adhesion and no falling.
And (3) sensitivity detection: blank control solution: the phosphate buffer solution containing bovine serum albumin was not detected to develop color. Detecting a sample solution containing 50mU/mLIAA standard: only the quality control line is colored, and the detection line is not colored. Detecting a sample solution containing 150mU/mLIAA standard: the quality control line and the detection line are colored. Detecting a sample solution containing 300mU/mLIAA standard: the quality control line and the detection line are colored.
And (3) specific detection: does not cross-react with other autoantibodies.
And (3) detecting the precision in the batch: detecting a sample solution containing 50mU/mLIAA standard: the reaction results are consistent, and the color development is uniform. Detecting a sample solution containing 150mU/mLIAA standard: the reaction results are consistent, and the color development is uniform. Detecting a sample solution containing 300mU/mLIAA standard: the reaction results are consistent, and the color development is uniform.
4. Quality standard of finished product
1.2 absorbance: 1ml of the sample to be detected is taken by a sample injector and put into a cuvette, the absorption value at the position of 395nm and 405nm of the wavelength is measured by a spectrophotometer, and the ratio of A395 to A405 is calculated to meet the requirement.
Example 3:
1.2 functional assays
And sticking the qualified large plate with the scratched film and the colloidal gold pad to be detected to prepare a test strip for functional detection.
1.2.1 sensitivity detection
1.2.1.1 reagents: blank control solution: 0.01mol/L Phosphate Buffer Solution (PBS) containing bovine serum albumin (BSA for short, the same applies below), pH7.2-pH7.4.
And (3) measuring a sample solution: sample solutions containing 50mU/mL, 150mU/mL, and 300mU/mL IAA standards.
The preparation method comprises the step of diluting IAA standard substance with BSA-containing phosphate buffer solution to obtain 50mU/mL, 150mU/mL and 300mU/mL respectively.
1.2.1.2 instruments: stopwatch
1.2.1.3 operating procedure:
1.2.1.3.1 extracting 3 products of the same batch, detecting blank control liquid, taking out the strip-shaped sample pad after one end is immersed in the sample liquid for at least 20 seconds, adding 3 drops of sample liquid into the sample adding hole if the strip-shaped sample pad is in a card shape, starting timing, reading the result within 10-15 minutes, judging the product to be qualified if the quality control line is uniformly colored and the detection line is not colored, otherwise judging the product to be unqualified;
1.2.1.3.250 mU/mL sample liquid detection amount and detection method are the same, the results of 3 product detection lines only develop color on the quality control line, the detection line does not develop color, the product is judged to be qualified, otherwise, the product is judged to be unqualified;
1.2.1.3.3150 mU/mL sample liquid detection amount and detection method are the same as those of 1.2.1.3.1, and if the result quality control line and the detection line of 3 product detection lines are colored, the product is judged to be qualified, otherwise, the product is judged to be unqualified;
1.2.1.3.3300 mU/mL sample liquid detection amount and detection method are the same as those of 1.2.1.3.1, and the result quality control line and detection line of 3 product detection lines are colored, so the product is judged to be qualified, otherwise, the product is judged to be unqualified.
1.2.2 specific assay
Does not cross-react with other autoantibodies.
1.2.3 determination of in-batch precision
1.2.3.1 reagents
And (3) measuring a sample solution: sample solutions containing 50mU/mL, 150mU/mL, and 300mU/mL IAA standards.
The preparation method comprises the steps of preparing 50mU/mL, 150mU/mL and 300mU/mL IAA samples by using a phosphate buffer solution containing BSA
1.2.3.2 instruments: stopwatch
1.2.3.3 working procedure
1.2.3.3.1 extracting 10 products of the same batch, detecting 50mU/mL IAA standard sample liquid, immersing one end of the strip-shaped sample pad into the sample liquid for at least 20 seconds, taking out, adding 3 drops of sample liquid into the sample adding hole if the sample pad is card-shaped, laying flat on a plane, starting timing, reading the result within 10-15 minutes, wherein the reaction result should be consistent, and the color should be uniform.
1.2.3.3.2150 mU/mL IAA standards were assayed as above.
1.2.3.3.3300 mU/mL IAA standards were assayed as above.
3) Method and control for inspecting gold pad
1.1 appearance: the appearance of the product is required to be observed by naked eyes.
1.2 functional assays
And sticking the qualified large plate with the scratched film and the colloidal gold pad to be detected to prepare a test strip for functional detection.
1.2.1 sensitivity detection
1.2.1.1 reagents: blank control solution: 0.01mol/L Phosphate Buffer Solution (PBS) containing bovine serum albumin (BSA for short, the same applies below), pH7.2-pH7.4.
And (3) measuring a sample solution: sample solutions containing 50mU/mL, 150mU/mL, and 300mU/mL IAA standards.
The preparation method comprises the step of diluting IAA standard substance with BSA-containing phosphate buffer solution to obtain 50mU/mL, 150mU/mL and 300mU/mL respectively.
1.2.1.2 instruments: stopwatch
1.2.1.3 operating procedure:
1.2.1.3.1 extracting 3 products of the same batch, detecting blank control liquid, taking out the strip-shaped sample pad after one end is immersed in the sample liquid for at least 20 seconds, adding 3 drops of sample liquid into the sample adding hole if the strip-shaped sample pad is in a card shape, starting timing, reading the result within 10-15 minutes, judging the product to be qualified if the quality control line is uniformly colored and the detection line is not colored, otherwise judging the product to be unqualified;
1.2.1.3.250 mU/mL sample liquid detection amount and detection method are the same, the results of 3 product detection lines only develop color on the quality control line, the detection line does not develop color, the product is judged to be qualified, otherwise, the product is judged to be unqualified;
1.2.1.3.3150 mU/mL sample liquid detection amount and detection method are the same as those of 1.2.1.3.1, and if the result quality control line and the detection line of 3 product detection lines are colored, the product is judged to be qualified, otherwise, the product is judged to be unqualified;
1.2.1.3.3300 mU/mL sample liquid detection amount and detection method are the same as those of 1.2.1.3.1, and the result quality control line and detection line of 3 product detection lines are colored, so the product is judged to be qualified, otherwise, the product is judged to be unqualified.
1.2.2 specific assay
Does not cross-react with other autoantibodies.
1.2.3 determination of in-batch precision
1.2.3.1 reagents
And (3) measuring a sample solution: sample solutions containing 50mU/mL, 150mU/mL, and 300mU/mL IAA standards.
The preparation method comprises the steps of preparing 50mU/mL, 150mU/mL and 300mU/mL IAA samples by using a phosphate buffer solution containing BSA
1.2.3.2 instruments: stopwatch
1.2.3.3 working procedure
1.2.3.3.1 extracting 10 products of the same batch, detecting 50mU/mL IAA standard sample liquid, immersing one end of the strip-shaped sample pad into the sample liquid for at least 20 seconds, taking out, adding 3 drops of sample liquid into the sample adding hole if the sample pad is card-shaped, laying flat on a plane, starting timing, reading the result within 10-15 minutes, wherein the reaction result should be consistent, and the color should be uniform.
1.2.3.3.2150 mU/mL IAA standards were assayed as above.
1.2.3.3.3300 mU/mLIAA standard sample solution determination method is the same as above.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (7)
1. The test strip for detecting the insulin-resistant autoantibody comprises a PVC base plate, a sample pad, a combination release pad, a nitrocellulose membrane and an absorption pad, wherein the sample pad, the combination release pad, the nitrocellulose membrane and the absorption pad are sequentially lapped on the PVC base plate.
2. The test strip of claim 1, wherein the anti-human IgG is murine.
3. The test strip of claim 2, wherein the second antibody is a goat anti-mouse secondary antibody.
4. A kit for detecting an anti-insulin autoantibody, comprising the test strip of any one of claims 1 to 3.
5. A method for preparing the test strip of any one of claims 1-3, comprising the steps of:
(1) preparing a colloidal gold solution;
(2) carrying out colloidal gold labeling on the anti-human IgG antibody; mixing the colloidal gold and the anti-human IgG antibody, centrifuging, and carrying out heavy suspension precipitation by using a PBS (phosphate buffer solution) solution to obtain an anti-human IgG-colloidal gold compound;
(3) preparing a gold-labeled combined release pad: uniformly spraying an anti-human IgG-colloidal gold compound on the combined release pad;
(4) preparing a nitrocellulose membrane: respectively coating the anti-human IgG secondary antibody and the insulin antigen on a quality control line and a detection line on a nitrocellulose membrane;
(5) assembling the test strip: and adhering the sample pad, the combined release pad, the coated nitrocellulose membrane and the absorption pad on a PVC plate in a dry environment, and cutting to obtain the test strip.
6. The method of claim 4, wherein the anti-human IgG is of murine origin.
7. The method according to claim 5, wherein the anti-human IgG secondary antibody is a goat anti-mouse secondary antibody.
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| CN202631539U (en) * | 2012-06-19 | 2012-12-26 | 中国人民解放军第三军医大学第一附属医院 | 1-type diabetes auto-antibody spectrum detection test paper |
| CN109946457A (en) * | 2019-03-04 | 2019-06-28 | 武汉科前生物股份有限公司 | A kind of colloidal gold strip detecting Porcine epidemic diarrhea virus |
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2019
- 2019-12-26 CN CN201911362124.3A patent/CN111103280A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1351655A (en) * | 1999-03-10 | 2002-05-29 | 马格福斯应用有限公司 | Method for cultivating cancer cells from human tissue and device for preparing tissue samples |
| CN101639480A (en) * | 2009-08-31 | 2010-02-03 | 杭州艾力康医药科技有限公司 | Test strip for quickly detecting antibodies in saliva for determining herpes simplex viruses |
| CN102621308A (en) * | 2011-01-28 | 2012-08-01 | 上海科新生物技术股份有限公司 | Colloidal gold chromatography anti-Jo-1 antibody detection test paper and preparation method thereof |
| CN202631539U (en) * | 2012-06-19 | 2012-12-26 | 中国人民解放军第三军医大学第一附属医院 | 1-type diabetes auto-antibody spectrum detection test paper |
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