CN102226811B - A preparation method of diagnostic test paper for detecting cardiac fatty acid binding protein in urine - Google Patents

Abstract

The invention relates to a diagnostic test paper for detecting an H-FABP. The test paper is characterized in that: the test paper is formed through adhering a sample pad, a cellulose nitrate membrane and an absorption pad to a base plate in sequence, arranging a gold-marking binding pad on the sample pad, and covering a PE label protective membrane on an upper layer; the gold-marking binding pad is coated with a labeled H-FABP mouse mono-antibody with colloidal gold; and the cellulose nitrate membrane is coated with a detection line which comprises the labeled H-FABP mouse mono-antibody and aquality control C line which comprises an H-FABP secondary antibody respectively. The diagnostic test paper has the advantages of strong specificity, high sensitivity and no need of any auxiliary instrument, and has a noninvasive advantage because a patient self can conveniently take a sample for the needed detection sample is urine; and simultaneously detection only needs 5 min, so the test paper provides a convenient and fast method for determining or eliminating myocardial infarction, thereby precious time is saved for early treatment.

Description

A preparation method of diagnostic test paper for detecting cardiac fatty acid binding protein in urine

technical field

The invention belongs to the technical field of immunochemical detection, and in particular relates to a preparation method of a diagnostic test paper for rapidly detecting cardiac fatty acid binding protein in urine by means of colloidal gold-labeled chromogenic immunochromatographic reaction.

Background technique

Acute myocardial infarction (AMI) is a common cardiovascular disease with high morbidity and mortality. The latest recommendations for the diagnosis of AMI are based on myocardial necrosis markers, combined with medical history, electrocardiogram, imaging changes, PCI, coronary angiography, and new diagnostic criteria for myocardial biochemical markers.

Heart-type fatty acid binding protein (heart-type fatty acid binding protein, H-FABP), as a marker of myocardial injury, has high sensitivity and specificity, is gradually being recognized by clinicians, and has become an effective indicator for early diagnosis of AMI.

H-FABP is a soluble cytoplasmic protein that exists specifically in myocardial tissue, with a molecular weight of only 14-15KD. Plasma and urine of normal people hardly contain H-FABP. When myocardial ischemia or damage occurs, H-FABP is quickly released into the blood, and is cleared from the kidneys and excreted in the urine; It can be seen that urine H FABP and plasma H FABP have the same diagnostic value for early acute myocardial infarction (AMI).

At present, the means of clinical detection of H-FABP, all use serum or plasma as the detection sample, using double antibody sandwich ELISA kit and other detection methods, the detection time is long, the blood sample needs to be separated and processed, and hemolysis is prone to occur, which cannot meet the requirements of determination or The requirement to provide rapid results for myocardial infarction is excluded.

Contents of the invention

In view of the deficiencies in the prior art, the object of the present invention is to provide a diagnostic test paper for detecting cardiac fatty acid binding protein in urine and a preparation method thereof through the research on detection technology of cardiac fatty acid binding protein. The diagnostic test paper not only has strong specificity and high sensitivity, and does not require any auxiliary equipment, but also the required test sample is urine, which is convenient for patients to take samples by themselves, and has the advantage of non-invasiveness; at the same time, the test time is only 5 minutes Excluding myocardial infarction provides a convenient and quick method, which saves precious time for early treatment.

The purpose of the present invention is achieved in this way: a diagnostic test paper for detecting cardiac fatty acid binding protein in urine, characterized in that: sample pad, nitrocellulose membrane, and absorbent pad are adhered to the base plate in sequence, and the gold standard binding pad is placed on the The sample pad is assembled by covering the upper layer with a PE label protective film; the gold-labeled binding pad is coated with colloidal gold-labeled H-FABP mouse monoclonal antibody; the nitrocellulose membrane is coated with H-FABP mouse monoclonal antibody The detection line T line and the quality control line C line coated with H-FABP secondary antibody.

The object of the present invention can also be achieved like this: the preparation method of the H-FABP mouse monoclonal antibody coated with colloidal gold label on the described gold label binding pad is as follows:

The preparation method of the colloidal gold-labeled H-FABP mouse monoclonal antibody coated on the gold-labeled binding pad is as follows:

(1) Take the colloidal gold solution, adjust the pH to 8.0-9.0 with 0.1M potassium carbonate solution, and stir for 5 minutes; add H-FABP mouse monoclonal antibody dropwise, continue stirring for 30 minutes, add 5% bovine serum albumin to make it final The concentration is 1%, stirred for 15 minutes, after standing still for 1 hour, stored at 4°C for later use;

(2) Centrifuge the gold-labeled H-FABP antibody conjugate prepared in the above step (1) at 12000 rpm and 4°C for 30 min, discard the supernatant, and use PB solution containing 1% BSA and 0.02% _NaN3 for the precipitate , resuspend the pellet to 1/10 of the original volume, store at 4°C, spray the gold-labeled H-FABP antibody conjugate on the glass fiber membrane, and dry at room temperature.

(3) Spray the gold-labeled H-FABP antibody conjugate prepared in the above (2) step on the glass fiber membrane, and after drying at room temperature, soak the membrane 3 times with 0.05mol/L TBS at pH 8.0-9.0, each Rinse once with deionized water for 3 minutes, immerse in 0.05mol/L citric acid buffer solution with pH3.0-5.0 for 5min, immerse in 37℃ silver staining solution for 20-30min, take out the membrane, rinse with deionized water, put Into the fixer for 1 min, rinse with deionized water, and air dry.

The detection line T line composed of H-FABP mouse monoclonal antibody coated on the nitrocellulose membrane is sprayed with H-FABP mouse monoclonal antibody on the nitrocellulose membrane, and dried at room temperature as the detection line T line.

The quality control line C line composed of the H-FABP secondary antibody coated on the nitrocellulose membrane is sprayed with the H-FABP secondary antibody on the nitrocellulose membrane and dried at room temperature as the quality control line C line.

The base plate is a PVC plate.

The present invention is a method for preparing diagnostic test paper for detecting cardiac fatty acid binding protein in urine, characterized in that:

(1) Preparation of gold-labeled binding pad:

1) Preparation of colloidal gold particles: Take 100ml of 0.01% HAuCl ₄ aqueous solution, add 2ml of 1% trisodium citrate aqueous solution, heat and boil for 15min-30min, until the color turns red; after cooling at room temperature, add 0.1Mol/L K ₂ CO ₃ 0.5 ml, mix well to obtain colloidal gold particles with a diameter of about 15nm, and store at 4°C;

2) Determination of the dosage ratio of colloidal gold and H-FABP mouse monoclonal antibody

a. Take 10ml of colloidal gold solution, adjust the pH to 8.0-9.0 with 0.1M potassium carbonate solution, stir for 5 minutes, and divide into 10 tubes, each with 1ml;

b. Serially dilute H-FABP mouse monoclonal antibody with 0.005Mol/L pH9.0 borate buffer solution to 5μg/ml-50μg/ml, take 1ml respectively, add to the above colloidal gold solution, and mix well; control Only add 1ml diluent to the tube;

c. After 5 minutes, add 0.1ml of 10% NaCl solution to each of the above tubes, mix well and let stand for 2 hours to observe the results;

d. Observation of the results, the control tube and the tubes in which the amount of H-FABP mouse monoclonal antibody was not enough to stabilize the colloidal gold showed coagulation from red to blue; while the amount of H-FABP mouse monoclonal antibody was added to reach Or each tube that exceeds the minimum stable amount remains red; the lowest dosage of H-FABP mouse monoclonal antibody that stabilizes the red color of 1ml colloidal gold solution is the minimum dosage of H-FABP mouse monoclonal antibody;

3) Preparation and purification of colloidal gold-labeled H-FABP antibody

a. Take 10ml colloidal gold solution, adjust pH to 8.0-9.0 with 0.1M potassium carbonate solution, stir for 5min; add H-FABP mouse monoclonal antibody drop by drop and continue stirring for 30min, add 5% bovine serum albumin (BSA) to make The final concentration is 1%, stirred for 15 minutes, after standing still for 1 hour, stored at 4°C for later use;

b. Purification: centrifuge the above colloidal gold H-FABP antibody conjugate at 12000 rpm and 4°C for 30 min. Discard the supernatant, and resuspend the precipitate in PB solution containing 1% BSA and 0.02% NaN ₃ to 1/10 of the original volume, and store it at 4°C for later use;

4) Treatment of silver staining: Spray the purified gold-labeled H-FABP mouse antibody conjugate on the glass fiber membrane, after drying at room temperature, soak the membrane 3 times with 0.05mol/L PH7.0-9.0TBS, each Rinse once with deionized water, immerse in 0.05mol/L PH3.0-5.0 citric acid buffer for 5min, immerse in silver staining solution at 37°C for 20-30min, take out the membrane, rinse with deionized water, put Into the fixer for 1 min, rinse with deionized water, air-dry, and store at 4°C for later use; the gold-labeled binding pad (2) is obtained;

(2) Preparation of nitrocellulose membrane coated with detection line T line and quality control line C line:

Spray the H-FABP mouse monoclonal antibody and the H-FABP secondary antibody on the nitrocellulose membrane, dry at room temperature, as the detection line T line and the quality control line C line, seal it, and store it at 4°C for later use;

(3) Assembly of the test paper: the bottom plate is assembled by pasting sample pads, nitrocellulose membranes, absorbent pads, and gold label binding pads on the sample pads from one end, and then covering the upper layer with a PE label protective film.

Compared with the existing detection method of H-FABP, the diagnostic test strip for detection of cardiac fatty acid binding protein in urine according to the present invention has the following advantages and significant progress:

(1) The test sample does not need to be processed: urine is used as the test sample. The urine H-FABP content and plasma H-FABP content have the same diagnostic value for early acute myocardial infarction (AMI). Patients can collect urine by themselves, and the sample It can be tested directly without pre-treatment, omitting the process of processing blood samples with serum or plasma as samples, and avoiding the problem of unusable samples caused by hemolysis that may occur during sample processing. It is convenient and non-invasive. advantage;

(2) Fast detection speed: the detection time only takes 5 minutes, which can quickly provide judgment basis for diagnosis;

(3) detection accuracy rate is high, specificity is strong: adopt H-FABP test paper made by the inventive method and H-FABP ELISA kit to carry out parallel detection respectively to 113 routine clinical urine samples, contrast detection result shows, the test paper detection result The coincidence rate was 97.35%, and the specificity was 96.3%. The test results are shown in Table 1

Test results Table 1

Description of drawings

Figure 1: Schematic diagram of the structure of the heart-type fatty acid binding protein (H-FABP) test strip

1-sample pad 2-gold standard binding pad 3-nitrocellulose membrane 4-absorbent pad 5-bottom plate 6-test line T line 7-quality control line C line

Figure 2: Schematic diagram of determination of test strip results for heart-type fatty acid binding protein (H-FABP)

A Negative B Positive C Invalid

Detailed ways

The principle that the present invention is based on is: human urine diffuses through the filter membrane of the antibody coated with red gold particles, combines with the coated antibody in the filter membrane, and in the T line region of the detection line, the H-FABP antigen-antibody compound The object begins to contact with the antibody coated in the T line area of the detection line, if the concentration of H-FABP in the urine is greater than 10ng/ml, the antigen-antibody complex in the T line area of the detection line is captured by the H-FABP monoclonal antibody, and A visible red band appears. The higher the concentration of H-FABP, the faster and darker the color band appears in the T line area of the detection line. The antibody coated with red gold particles diffuses to the C-line area of the quality control line and is captured by the H-FABP secondary antibody to form a red band in the quality control area.

The invention relates to a diagnostic test strip for detecting cardiac fatty acid binding protein in urine, which consists of a sample pad 1, a gold standard binding pad 2, a nitrocellulose membrane 3, an absorption pad 4, a bottom plate 5, and a detection line T line 6 And quality control line C line 7 composition (see Figure 1).

A method for preparing a diagnostic test strip for detecting cardiac fatty acid binding protein in urine, which the present invention relates to, comprises the following steps:

1. Preparation of colloidal gold solution and gold-labeled antibody: Prepare colloidal gold solution by trisodium citrate reduction method, determine the ratio of colloidal gold and H-FABP antibody dosage, prepare gold-labeled H-FABP antibody marker, and use low-temperature ultracentrifugation Gold-labeled antibody was purified by the method;

2. Treatment of silver staining: Spray the gold-labeled H-FABP antibody on the glass fiber membrane, then stain with silver staining solution, fix and dry; the color signal can be further amplified by strengthening the immunogold staining with silver, and the other The sensitivity can be equivalent to that of enzyme-linked immunosorbent technology, thereby improving the sensitivity and accuracy of visual judgment results.

3. Preparation of test line T line and quality control line C line: H-FABP mouse monoclonal antibody and H-FABP secondary antibody were sprayed on nitrocellulose membrane respectively, dried, and used as test line T line and quality control line C line ; The secondary antibody refers to an antibody that can bind to an antibody, that is, an antibody of an antibody, and its main function is to detect the presence of an antibody. The "H-FABP secondary antibody" can be a rabbit anti-mouse H-FABP monoclonal antibody or a goat anti-mouse H-FABP monoclonal antibody. The present invention uses a rabbit anti-mouse H-FABP monoclonal antibody.

4. Assembling test strips: Paste, assemble, cut strips, and seal the processed nitrocellulose membrane, glass fiber membrane, sample pad, absorbent pad and other materials in sequence, and store at 4°C.

The method for using a diagnostic test paper for detecting cardiac fatty acid binding protein in urine is as follows:

1. Insert the test strip vertically into the urine sample cup, be careful not to exceed the sample pad, take it out after at least 3 seconds and place it flat on a clean non-absorbent material plane, and observe the result for 5 minutes.

2. Judgment of results (see Figure 2):

1) Negative: A red band appears on the C line of the quality control line, but no red band appears on the T line of the test line, indicating that there is no H-FABP in the test sample;

2) Positive: a red band appears at the T line of the test line and the C line of the quality control line, and it is judged as positive;

3) Invalid: Only when the color develops on the T line of the test line or no obvious band appears on the T line of the test line and the C line of the quality control line, the test strip test is considered invalid.

The color bands of the C line of the quality control line and the T line of the detection line may appear dark or light depending on the amount of H-FABP in the urine. When the concentration of H-FABP is high, the T line of the detection line is very obvious, and the C line of the quality control line may change Very weak, normal result.

Test strip specificity test of the present invention:

Troponin, C-reactive protein standard solution (concentration is 20ng/ml), pregnant woman urine and H-FABP standard solution (20ng/ml) are detected with test strip of the present invention, only H-FABP standard solution appears as a result Obvious detection line and quality control line, the rest of the samples were tested negative. It shows that the test strip has good specificity.

Test strip sensitivity test of the present invention:

After the H-FABP antigen is diluted 1:100 times, it is diluted by 10 times, and then each dilution is detected with the test strip of the present invention. The results show that the minimum detection amount is 1:1000, that is, 10ng/ml .

Example

1. Preparation of colloidal gold particles: Take 100ml of 0.01% HAuCl ₄ aqueous solution, add 2ml of 1% trisodium citrate aqueous solution, heat and boil for 15min-30min, until the color turns red. After cooling at room temperature, add 0.1Mol/L K ₂ CO ₃ 0.5ml, mix well to obtain colloidal gold particles with a diameter of about 15nm, and store at 4°C.

2. Determination of the dosage ratio of colloidal gold and H-FABP mouse monoclonal antibody

(1) Take 10ml of colloidal gold solution, adjust the pH to 8.0-9.0 with 0.1M potassium carbonate solution, stir for 5min, and divide into 10 tubes, each with 1ml.

(2) Serially dilute H-FABP mouse monoclonal antibody with 0.005Mol/L pH9.0 borate buffer solution to 5μg/ml~50μg/ml, take 1ml respectively, add to the above colloidal gold solution, and mix well. Only add 1ml diluent to the control tube.

(3) After 5 minutes, add 0.1ml of 10% NaCl solution to each of the above tubes, mix well and let stand for 2 hours to observe the results.

(4) Observation of the results, the control tube and the tubes in which the amount of H-FABP antibody added was not enough to stabilize the colloidal gold showed coagulation from red to blue; while the amount of H-FABP antibody added reached or exceeded the minimum stable amount The tubes remain red unchanged. The minimum dosage of H-FABP antibody is the minimum dosage of H-FABP antibody that stabilizes the red color of 1ml colloidal gold solution. In actual work, it can be appropriately increased by 10% to 20%.

3. Preparation and purification of colloidal gold-labeled H-FABP antibody

①Take 10ml of colloidal gold solution, adjust the pH to 8.0-9.0 with 0.1M potassium carbonate solution, and stir for 5 minutes; add H-FABP mouse monoclonal antibody drop by drop and continue stirring for 30 minutes, add 5% bovine serum albumin (BSA) to make it The final concentration was 1%, stirred for 15 minutes, and after standing still for 1 hour, it was stored in a 4°C refrigerator for later use.

② Purification: Centrifuge the above colloidal gold H-FABP antibody conjugate at 12000 rpm and 4°C for 30 min. Discard the supernatant, and use the PB solution containing 1% BSA (containing 0.02% NaN ₃ ) for the precipitate. The PB solution is phosphate buffer (phosphate buffer solution), resuspend the precipitate to 1/10 of the original volume, and store it at 4°C for later use .

4. Treatment of silver staining: Spray the gold-labeled H-FABP antibody on the glass fiber membrane, and after drying at room temperature, soak the membrane 3 times with 0.05mol/lPH7.0-9.0tbs (TBS refers to Tris buffered saline (TrisBuffered saline) , Saline, TBS) Tris is the English name of Tris), each time for 3 minutes, washed once with deionized water, immersed in 0.05mol/l pH3.0-5.0 citric acid buffer for 5 minutes, and immersed in 37 ° C 20-30min in silver staining solution. Take the film out, rinse it with deionized water, put it in the fixer solution for 1 min, rinse it with deionized water, and air dry. Strengthening immunogold staining with silver can further amplify the chromogenic signal, and its sensitivity can be comparable to that of enzyme-linked immunosorbent technology, thereby improving the sensitivity and accuracy of visual judgment results.

5. Preparation of detection line T line and quality control line C line: Spray H-FABP mouse monoclonal antibody and H-FABP secondary antibody on nitrocellulose membrane respectively, and dry at room temperature as the detection line T line and quality control line Line C, sealed, and stored at 4°C for later use.

6. Assembling test strips: Paste, assemble, cut strips, and seal the processed nitrocellulose membrane, glass fiber membrane, sample pad, absorbent pad and other materials in sequence, and store at 4°C. See attached drawing 1.

Sample pad 1: Placed at the front end of the test strip for sample injection, control the flow rate of the test solution to ensure that the test solution flows into the gold-labeled binding pad at a uniform speed, so that the gold-labeled antibody is specific to the H-FABP in the test solution sexually combined. The material used is glass fiber membrane.

Gold label binding pad 2: It is the carrier of colloidal gold markers. When the test solution flows through the gold label binding pad through the sample pad, the colloidal gold conjugate is released from the gold label binding pad and is specific to H-FABP in the test solution. Combined to form an immune complex, and then flow to the nitrocellulose membrane together with the test solution. The selected material is glass fiber membrane.

Nitrocellulose membrane 3 (NC membrane): It is the display area of the final result, generally there are two lines, the detection line T line and the quality control line C line.

Absorbent pad 4: placed at the end of the test strip, used to control the absorption of the test solution, increase the amount of test solution flowing into the test strip, to wash away the free colloidal gold conjugates on the NC membrane, thereby reducing the background Interference has increased sensitivity. The material used is plant fiber paper.

Bottom plate 5: It has the function of carrying sample pad, gold standard binding pad, NC film and absorbing pad to form a real strip shape, which is convenient for test operation. The selected material is PVC board, which is coated with pressure-sensitive adhesive that is inert to biologically active molecules.

Detection line T line 6: spray H-FABP mouse monoclonal antibody on the nitrocellulose membrane as a detection line for detecting H-FABP antigen in the test solution.

Quality control line C line 7: Spray H-FABP secondary antibody on nitrocellulose membrane as quality control line for detection of colloidal gold conjugates.

Paste the sample pad 1, the gold label binding pad 2, the nitrocellulose membrane 3, the absorption pad 4 on the bottom plate 5 in sequence, the H-FABP antibody-colloidal gold marker is coated on the gold label binding pad 2, and the nitrocellulose membrane is sprayed H-FABP mouse monoclonal antibody is used as the detection line T line and H-FABP secondary antibody is used as the quality control line C line. The specific production method: put the gold label binding pad on the sample pad, the sample pad is glued to the bottom plate, the nitrocellulose membrane 3 (NC membrane) is pasted on the bottom plate 5 , and the absorption pad 4 is also pasted on the bottom plate 5 . It is assembled by staggering each other by 2mm from left to right and pasting on the bottom plate 5, and then covering the upper layer with a PE label protective film. Cut into 2-4mm wide test strips and store at 4°C.

Claims (3)

1. A preparation method for detecting cardiac fatty acid binding protein diagnostic test paper in urine, characterized in that: (1) Preparation of gold-labeled binding pad (2): 1) Preparation of colloidal gold particles: Take 100ml of 0.01% HAuCl ₄ aqueous solution, add 2ml of 1% trisodium citrate aqueous solution, heat and boil for 15min-30min, until the color turns red; after cooling at room temperature, add 0.1mol/L K ₂ CO ₃ 0.5 ml, mix well to obtain colloidal gold particles with a diameter of about 15nm, and store at 4°C; 2) Determination of the dosage ratio of colloidal gold and H-FABP mouse monoclonal antibody a. Take 10ml of colloidal gold solution, adjust the pH to 8.0-9.0 with 0.1M potassium carbonate solution, stir for 5 minutes, and divide into 10 tubes, each with 1ml; b. Serially dilute the H-FABP mouse monoclonal antibody with 0.005mol/L pH9.0 borate buffer solution to 5μg/ml-50μg/ml, take 1ml respectively, add to the above colloidal gold solution, and mix well; Only add 1ml diluent to the tube; c. After 5 minutes, add 0.1ml of 10% NaCl solution to each of the above tubes, mix well and let stand for 2 hours to observe the results; d. Observation of the results, the control tube and the tubes in which the amount of H-FABP mouse monoclonal antibody was not enough to stabilize the colloidal gold showed coagulation from red to blue; while the amount of H-FABP mouse monoclonal antibody was added to reach Or each tube that exceeds the minimum stable amount remains red; the lowest dosage of H-FABP mouse monoclonal antibody that stabilizes the red color of 1ml colloidal gold solution is the minimum dosage of H-FABP mouse monoclonal antibody; 3) Preparation and purification of colloidal gold-labeled H-FABP antibody a. Take 10ml colloidal gold solution, adjust the pH to 8.0-9.0 with 0.1M potassium carbonate solution, and stir for 5 minutes; add H-FABP mouse monoclonal antibody drop by drop and continue stirring for 30 minutes, add 5% bovine serum albumin (BSA) to make The final concentration is 1%, stirred for 15 minutes, after standing still for 1 hour, stored at 4°C for later use; b. Purification: centrifuge the above colloidal gold H-FABP antibody conjugate at 12000 rpm and 4°C for 30 min. Discard the supernatant, and resuspend the precipitate in PB solution containing 1% BSA and 0.02% NaN ₃ to 1/10 of the original volume, and store at 4°C for later use; 4) Treatment of silver staining: Spray the purified gold-labeled H-FABP antibody conjugate on the glass fiber membrane, after drying at room temperature, soak the membrane with 0.05mol/L pH7.0-9.0TBS for 3 times, each time for 3min , Rinse once with deionized water, immerse in 0.05mol/L pH3.0-5.0 citric acid buffer for 5 minutes, immerse in silver staining solution at 37°C for 20-30 minutes, take out the membrane, rinse with deionized water, and put it in the fixer solution for 1 min, rinsed with deionized water, air-dried, and stored at 4°C for later use; the gold-labeled binding pad (2) was obtained; (2) Preparation of nitrocellulose membrane (3) coated with detection line T line (6) and quality control line C line (7): Spray H-FABP mouse monoclonal antibody and H-FABP secondary antibody on nitrocellulose membrane (3), dry at room temperature, as detection line T line (6) and quality control line C line (7), seal, 4 Store at ℃ for later use; (3) Assembling the test paper: stick the sample pad (1), nitrocellulose membrane (3), absorbent pad (4) and gold standard binding pad (2) on the sample pad (1) on the bottom plate (5) in sequence from one end , and then assembled by covering the PE label protective film on the upper layer. 2. A diagnostic test paper for detecting cardiac fatty acid binding protein in urine made by the preparation method of claim 1, characterized in that: sample pad (1), nitrocellulose membrane (3), and absorbent pad (4) in sequence Adhered to the bottom plate (5), the gold label binding pad (2) is placed on the sample pad (1), and the upper layer is covered with a PE label protective film to assemble; the gold label binding pad (2) is coated with colloidal gold Labeled H-FABP mouse monoclonal antibody; the detection line T line (6) composed of H-FABP mouse monoclonal antibody coated on the nitrocellulose membrane (3) and the matrix coated with H-FABP secondary antibody Control Line C Line (7). 3. The diagnostic test paper for detecting cardiac fatty acid binding protein in urine according to claim 2, characterized in that: the bottom plate (5) is a PVC plate.

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