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

Active Publication Date: 2011-10-26
JILIN JISHENG PHARMA
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Problems solved by technology

[0005] 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, t

Method used

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  • Preparation method of diagnostic test paper for detecting H-FABP (heart-type fatty acid binding protein)
  • Preparation method of diagnostic test paper for detecting H-FABP (heart-type fatty acid binding protein)
  • Preparation method of diagnostic test paper for detecting H-FABP (heart-type fatty acid binding protein)

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preparation example Construction

[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...

Embodiment

[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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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

technical field [0001] 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 [0002] 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. [0003] 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. [0...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/558G01N33/577G01N33/544G01N33/532
Inventor 郭超王丽丽
Owner JILIN JISHENG PHARMA
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