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Colloidal gold test paper strip for semiquantitatively detecting concentration of GFAP (Glial Fibrillary Acidic Protein) in human serum

A technology of semi-quantitative detection and colloidal gold test paper, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of shortened detection time, cumbersome detection methods, long detection time, etc., and achieve low cost, high specificity, and high sensitivity Effect

Active Publication Date: 2012-02-15
山东莱博生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the detection of GFAP in clinical diagnosis is mainly based on the ELISA experimental method, which has been widely used in the past two decades because of its high specificity and sensitivity, but at the same time, the detection method of this method is cumbersome and requires a long test time (1 ~2 hours), and it is often necessary to accumulate a certain amount of samples before testing can be carried out, and the detection is delayed. However, the colloidal gold method has many advantages and can make up for its shortcomings: a) Fast: the entire detection process only takes 5-10 minutes, Significantly reduced detection time
However, the search results show that there is no report on the use of colloidal gold method to detect the concentration of GFAP in human serum

Method used

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  • Colloidal gold test paper strip for semiquantitatively detecting concentration of GFAP (Glial Fibrillary Acidic Protein) in human serum
  • Colloidal gold test paper strip for semiquantitatively detecting concentration of GFAP (Glial Fibrillary Acidic Protein) in human serum
  • Colloidal gold test paper strip for semiquantitatively detecting concentration of GFAP (Glial Fibrillary Acidic Protein) in human serum

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Step 1: Preparation of detection diaphragm

[0031] Dilute the anti-GFAP capture monoclonal antibody to four different concentrations with PBS buffer, and spot them on the nitrocellulose membrane with a spotter from high to low. Line spacing 5mm. Its coating amount represents that it can detect 2ng / ml, 4ng / ml, 8ng / ml and 16ng / ml. In the same way, 200 μg / ml of goat anti-mouse IgG was used as the quality control line after the detection line with a spotter, the line width was 1-1.5 mm, and the distance from the nearest detection line was 8 mm. Blocked with PBS buffer containing 1% BSA for 2 hours, dried in the shade at room temperature, and stored at 4°C.

[0032] Step 2: Preparation of Colloidal Gold Bonded Membrane

[0033] 1) Anti-GFAP antibody

[0034] Monoclonal antibodies were purchased from Santa Cruz Biotechnology.

[0035] 2) Preparation of anti-GFAP antibody-colloidal gold complexes

[0036] Colloidal gold with a diameter of 20 nm was prepared by trisodium c...

Embodiment 2

[0043] Application of colloidal gold test strip for semi-quantitative detection of GFAP concentration in human serum

[0044] (1) Detection of GFAP concentration in human serum with test strips

[0045] Aspirate 120 μl of serum sample with a pipette, and drop it vertically on the sample adding place of the colloidal gold test strip. Start the timer, wait for the red band to appear, and compare it with the standard colorimetric card within 10 minutes. If it exceeds 20 minutes, the result will be invalid.

[0046] (2) Interpretation of results

[0047] The four concentration gradient marks (ie detection lines) involved from top to bottom in the colloidal gold test strip are: 16ng / ml, 8ng / ml, 4ng / ml, and 2ng / ml. The test results are explained as follows:

[0048] When only one line of the quality control line appears, the GFAP concentration is less than 2ng / ml.

[0049] When two lines containing the quality control line appear, the GFAP concentration is between 2ng / ml and 4ng...

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Abstract

The invention discloses a colloidal gold test paper strip for semiquantitatively detecting the concentration of a GFAP (Glial Fibrillary Acidic Protein) in a human serum. The colloidal gold test paper strip is produced by adhering a detection diaphragm, a colloidal gold combined diaphragm and a water-absorbing paper layer to a PVC (Poly Vinyl Chloride) board, wherein the colloidal gold combined diaphragm is coated by a colloidal gold-labeled antibody for resisting the GFAP; a detection line and a quality control line are arranged on the detection diaphragm; a capture antibody for resisting the GFAP is fixedly arranged on the detection line; a rabbit anti-mouse IgG (Immunoglobulin G) is fixedly arranged on the quality control line; and a sampling part, the colloidal gold combined diaphragm, the detection line, the quality control line and the water-absorbing paper are sequentially arranged on the test paper strip from the bottom up. The test paper strip disclosed by the invention can be used for semiquantitatively detecting the GFAP, has the advantages of high specificity, high sensitiveness, high accuracy, quickness, simpleness and convenience in operation, low expense, no need ofany apparatus, time-and-labor saving and the like as well as has favorable application prospect.

Description

technical field [0001] The invention relates to a colloidal gold test strip, in particular to a colloidal gold test strip for semi-quantitatively detecting the concentration of glial fibrillary acidic protein (GFAP) in human serum. Background technique [0002] Epidemiological research shows that cerebrovascular disease, heart disease and malignant tumor constitute the three major causes of human death. Compared with western developed countries, the morbidity and mortality of cerebrovascular disease in my country is significantly higher than that of cardiovascular disease. It is estimated that there are about 2 million new stroke patients in China every year; about 1.5 million people die of stroke every year; the number of surviving patients is 6 million to 7 million; and 70% to 80% of the survivors are left with paralysis and aphasia. and other serious disabilities, which bring a heavy burden to society and families. Among them, cerebral hemorrhage is a common and frequen...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N21/76
Inventor 王红王佳颖朱之炜郑祖惠丁兴龙韩娟张增丽李夫东王恒江长林
Owner 山东莱博生物科技有限公司
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