Preparation method of diagnostic test paper for detecting H-FABP (heart-type fatty acid binding protein)
A fatty acid combination and heart technology, applied in the field of immunochemical detection, can solve the problems of long detection time, inability to meet the requirements of rapid results of myocardial infarction, and prone to hemolysis, etc., to achieve fast detection speed, strong specificity, and accurate detection high rate effect
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[0043] 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:
[0044] 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;
[0045] 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.
[0046] 3. Prepa...
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[0060] 1. Preparation of colloidal gold particles: take 0.01% HAuCl 4 100ml of aqueous solution, add 2ml of 1% trisodium citrate aqueous solution, heat and boil for 15min to 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.
[0061] 2. Determination of the dosage ratio of colloidal gold and H-FABP mouse monoclonal antibody
[0062] (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.
[0063] (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.
[0064] (3) After 5 minutes, add 0.1ml of 10% NaCl solution to each of the above tubes, mix well and...
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