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

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

Active Publication Date: 2013-03-06
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 membrane

[0031] Dilute the anti-GFAP capture monoclonal antibody to four different concentrations with PBS buffer, and spot on the nitrocellulose membrane with a film spotter from high to low, each line is 20 μl, and the line width is 1-1.5 mm. The line spacing is 5mm. Its coating amount means that 2ng / ml, 4ng / ml, 8ng / ml, and 16ng / ml can be detected. In the same way, spot 200 μg / ml goat anti-mouse IgG after the test line as a quality control line with a film spotter, with a line width of 1-1.5 mm and a distance of 8 mm from the nearest test line. Block with PBS buffer containing 1% BSA for 2 hours, dry in the shade at room temperature, and store at 4°C.

[0032] Step 2 Preparation of Colloidal Gold Bonded Film

[0033] 1) Anti-GFAP antibody

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

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

[0036] Colloidal gold with a diameter of 20nm ...

Embodiment 2

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

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

[0045] Use a pipette to draw 120 μl of serum sample, and drop it vertically onto the sample loading site of the colloidal gold test strip. Start timing, wait for the red band to appear, and compare it with the standard color card within 10 minutes. If it exceeds 20 minutes, the result will be invalid.

[0046] (2) Interpretation of results

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

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

[0049] When two lines containing quality control lines appear, it means that the concentration of GFAP is between 2ng / ml a...

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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 of any 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 surveys show that cerebrovascular disease, heart disease, and malignant tumors constitute the three major causes of human death. Compared with developed countries in the West, the morbidity and mortality of cerebrovascular diseases in my country are significantly higher than those of cardiovascular diseases. It is estimated that there are about 2 million new stroke patients in China every year; about 1.5 million patients die of stroke every year; 6 million to 7 million surviving patients; and 70% to 80% of the survivors are left with paralysis and aphasia. Severe disabilities, such as disability, bring heavy burdens to society and families. Among them, cerebral hemorrhage is a common and frequently-o...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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