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Homo-type cysteine immunity colloidal gold detecting test paper and method for producing the same

A technology of homocysteine ​​and colloidal gold, which is applied in biological testing, measuring devices, material inspection products, etc., can solve the problems of undetected immunocolloidal gold chromatography rapid diagnostic test strips, etc., and achieve sensitivity, specificity, The effect of simple operation and low price

Inactive Publication Date: 2008-08-20
BEIJING HUAANFO BIOMEDICAL RES CENT +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are no immunocolloidal gold chromatography rapid diagnostic test strips for the detection of HCY at home and abroad, including literature reports and invention patents

Method used

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  • Homo-type cysteine immunity colloidal gold detecting test paper and method for producing the same
  • Homo-type cysteine immunity colloidal gold detecting test paper and method for producing the same
  • Homo-type cysteine immunity colloidal gold detecting test paper and method for producing the same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Homocysteine ​​(HCY) colloidal gold immune test strip preparation method, it comprises the steps:

[0040] 1. Preparation of SAH-BSA antigen:

[0041] Dissolve appropriate amount of BSA and SAH in 15ml PBS Erlenmeyer flasks respectively, stir with magnetic stirrer for 10 minutes, add SAH solution into BSA solution, stir while adding, then add 25% glutaraldehyde, react for 5 hours, gel layer The conjugate was separated by analysis, dialyzed in PBS for 24 hours, and concentrated.

[0042] 2. SAH monoclonal antibody preparation:

[0043] Balb / c mice were immunized with SAH-BSA conjugates. After multiple basic immunizations and booster immunizations, the splenocytes of the mice were collected, and the splenocytes were combined with Sp2 / 0 myeloma cells to prepare SAH mAb using PEG cell fusion technology.

[0044] Inject the hybridoma cells into the peritoneal cavity of the mouse, prepare ascites, and purify the monoclonal antibody by chromatography.

[0045] 3. Preparatio...

Embodiment 2

[0052] Homocysteine ​​(HCY) colloidal gold immune test strip preparation method, it comprises the steps:

[0053] SAH-BSA antigen and SAH monoclonal antibody were prepared in Example 1.

[0054] 1. Preparation of colloidal gold:

[0055] Measure a certain amount of lotion in a clean 250ml Erlenmeyer flask, boil for 5 minutes, change the pickling solution again, and boil for 5 minutes; measure a certain amount of ultrapure water in the flask, boil for 5 minutes, and change it again Ultrapure water, boiled for 5 minutes; discard the ultrapure water, weigh 200ml of ultrapure water in the flask, heat to boiling, add 10ml of 1% chloroauric acid, stir while cooking, heat to boiling; add 2 % trisodium citrate 7.5ml, stir rapidly under boiling water until the color of the chloroauric acid solution is slowly stable, and after being red, continue to boil for 9 minutes to make medium-sized colloidal gold particles.

[0056] 2. Colloidal gold probe labeling:

[0057] The SAH monoclonal...

Embodiment 3

[0061] Homocysteine ​​(HCY) colloidal gold immune test strip preparation method, it comprises the steps:

[0062] SAH-BSA antigen and SAH monoclonal antibody were prepared in Example 1.

[0063] 1. Preparation of colloidal gold:

[0064] Measure a certain amount of lotion in a clean 250ml Erlenmeyer flask, boil for 5 minutes, change the pickling solution again, and boil for 5 minutes; measure a certain amount of ultrapure water in the flask, boil for 5 minutes, and change it again Ultrapure water, boiled for 5 minutes; discard the ultrapure water, weigh 200ml of ultrapure water in the flask, heat to boiling, add 10ml of 1% chloroauric acid, stir while cooking, heat to boiling; add 2 % trisodium citrate 6.6ml, stir rapidly under boiling water, until the color of the chloroauric acid solution is slowly stable, after being red, continue to boil for 9 minutes to make larger colloidal gold particles.

[0065] 2. Colloidal gold probe labeling:

[0066] The SAH monoclonal antibody...

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Abstract

The invention provides a homocysteine immune colloidal gold test strip and the preparing method thereof. The test strip is strip shaped which comprises water uptake pad (2), nitrocellulose membrane (NC membrane) (3) with specific coating, capturing line (4) on NC film and quality control line (5), glass fiber colloidal gold combined pad (6) labeled by specific antibody with colloidal gold coating, and sample pad (7), which are subsequently combined on bottom plate (1), the glass fiber colloidal gold combined pad (6) is coated with colloidal gold specific antibody which is S adenosine homocysteine (SAH) monoclonal antibody. The test strip provided by the invention is used to can test homocysteine level in serum, and has the merits of easy operation, fast speed, sensitivity, specificity and so on, with good prospects.

Description

technical field [0001] The invention relates to a test strip for detecting HCY level in blood by colloidal gold immunochromatography technology and a preparation method thereof, belonging to the field of medical examination. Background technique [0002] Homocysteine ​​(homocysteine, Hcy) is an intermediate product of methionine metabolism in the human body, and also participates in the transsulfuration pathway in vivo. Hcy is remethylated to methionine via the folate cycle by B12 in the presence of 5-methyltetrahydrofolate, and the resulting 5,10-dimethyltetrahydrofolate is reduced to 5 by methyltetrahydrofolate reductase (MTHFR) -Methyltetrahydrofolate. Hcy can also be remethylated to methionine by glycine trimethylinyl salt. Remethylation is completed in the liver and kidney. In the methionine cycle, methionine in food is converted into S-adenosylmethionine (SAM). SAM is used as a substrate for the methyl group donor of methyltransferase. S-adenosylhomocysteine ​​(SAH)...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/577G01N33/558G01N33/544G01N33/532
Inventor 吴瑕王玉王燕徐希平于多张彦明
Owner BEIJING HUAANFO BIOMEDICAL RES CENT
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