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SAA (serum amyloid A) immunochromatographic test strip and preparation method and test method thereof

An immunochromatographic detection and test strip technology, which is applied in the field of preparation of immunochromatographic detection test strips, can solve the problems of not asking the market, and achieve the effects of short detection time, high specificity, and high sensitive detection performance.

Inactive Publication Date: 2016-06-01
SUZHOU DONGNI BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0017] In the diagnosis of autoimmune diseases, there have been many immunochromatographic test strips, but the immunochromatographic test strips based on SAA detection have not yet come into the market at home and abroad.

Method used

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  • SAA (serum amyloid A) immunochromatographic test strip and preparation method and test method thereof
  • SAA (serum amyloid A) immunochromatographic test strip and preparation method and test method thereof
  • SAA (serum amyloid A) immunochromatographic test strip and preparation method and test method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Preparation of each component of the gold standard test paper of embodiment 1

[0049] 1 Preparation of colloidal gold solution

[0050] 0.01% HAuCl 4 Heat the solution to boiling, quickly add every 100mLHAuCl 4 The solution is added with an appropriate amount of reducing agent solution, the color changes from blue, then light blue, blue, then red after heating, and transparent orange-red after boiling for 7-10 minutes. Then filter with ultrafiltration or microporous membrane (0.45uM) to remove polymers and other possible impurities. The prepared colloidal gold should be pure, translucent, free of sediment and floating matter, and discard when oily matter and a large amount of black granular precipitate impurities appear on the liquid surface.

[0051] Wherein the reducing agent used can be trisodium citrate, tannic acid-trisodium citrate, white phosphorus, preferably use trisodium citrate, more preferably use 1% trisodium citrate. The glass containers used therein ...

Embodiment 2

[0075] Example 2 Preparation of colloidal gold-labeled anti-SAA antibody protein detection test paper 1-3

[0076] The sample pads prepared in Example 1 and dried, the binding pads, the analysis membrane, and the absorbent paper were cut into 1.7cm, 0.8cm, 2.5cm, and 1.5cm wide narrow strips with a cutting machine, according to figure 1 The way is overlapped into large slabs, and the large slabs are cut into single servings with a strip cutter. The width of each serving varies according to the jamming of the bottom plate. The preferred width of the present invention is 3mm. Assemble the test paper that has been cut for a single person into the prepared test paper card, so that the sample loading window corresponds to the sample pad of the test paper, and the result display window corresponds to the detection area and quality control area. 20~30%.

[0077] The components of gold standard test strips 1~3 are as follows:

[0078]

Embodiment 3

[0079] Embodiment 3 sample processing

[0080] Serum sample: Take 1~5mL of whole blood in a serum collection tube, let it stand for 30min~2h, centrifuge at 3000~5000g for 5~10min, and take the supernatant. According to the detection accuracy of the test strip, dilute the sample 0-100 times with the sample buffer solution, take 50-100uL dropwise into the sample hole of the test strip, and observe the result after standing for 5-20min.

[0081] Plasma sample: Take 1~5mL of whole blood, mix it in a sodium citrate or sodium heparin anticoagulant tube, centrifuge at 1000~3000g for 5~10min, and take the supernatant to get the plasma sample. According to the detection accuracy of the test strip, dilute the sample 0-100 times with the sample buffer solution, take 50-100uL dropwise into the sample hole of the test strip, and observe the result after standing for 5-20min.

[0082] Whole blood sample: Take about 50uL of fresh blood from the fingertip or earlobe, drop it into the sample ho...

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Abstract

The invention discloses an SAA (serum amyloid A) immunochromatographic test strip and a preparation method and a test method thereof. The test strip comprises a sample pad, a combination pad, an analyzing membrane, a water-absorbing pad, a bottom board, a testing tape T line and a quality control tape C line, wherein the analyzing membrane is adhered on the upper portion of the bottom board, the combination pad and the water-absorbing pad are adhered at two ends of the upper portion of the analyzing membrane respectively, the sample pad is adhered at one end of the upper portion of the combination pad, and the testing tape T line and the quality control tape C line are adhered on the analyzing membrane. The SAA immunochromatographic test strip and the preparation method and the test method thereof have the advantages that anti-SAA antibody protein is used for achieving rapid testing of high sensitivity and high specificity of SAA antibodies in a sample by indirect immunoassay, so that bases for auxiliary diagnosis of diseases such as infectious diseases, tissue damage necrotic diseases, atherosclerotic diseases, immune diseases and rejection after organ transplantation are provided.

Description

[0001] The invention relates to the field of immunoassay detection. Specifically, the present invention relates to a preparation method and application of an immunochromatographic test paper for detecting SAA. Background technique [0002] Rosenthal et al. isolated a protein with a molecular weight of about 12,000 from serum in 1976, which has similar immunogenicity to the previously discovered tissue amyloid A with a molecular weight of 8,500, called serum amyloid A (serumamyloid A, SAA) ). SAA is the precursor of tissue amyloid A, which belongs to the acute phase protein, which increases during tissue injury and inflammatory response, and affects cell adhesion, migration, proliferation and aggregation. At present, acute phase protein (APP) can be divided into five categories, that is, APP involved in the inhibition of protease; APP involved in blood coagulation and fibrinolysis; APP belonging to complement components; APP involved in operation; other APPs include C-respons...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/68
Inventor 王峰王玉立卜天
Owner SUZHOU DONGNI BIOTECH CO LTD
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