Method of regenerating ELISA reaction plate for enzyme-link immunosorbent assay

An enzyme-linked immunosorbent assay and reaction plate technology, which can be used in biological testing, material inspection products, etc., and can solve problems such as weak short-range force

Inactive Publication Date: 2003-06-04
NANJING RED CROSS BLOOD CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to overcome the disadvantage that the above-mentioned reaction plate cannot be reused after being used once. According to the basic principle of ELISA, although the combination of antigen and antibody is very specific, it is not through ionic bond or covalent bond, but through a very weak bond. These forces are very weak under certain external conditions, so that the antigen and antibody complex dissociate, making the antigen-antibody Combination as a reversible chemical reaction

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0006] Example 1. Use the anti-HIV kit batch number 971113 produced by Xiamen Xinchuang. According to the operating instructions, after adding the substrate for color development, suck out the positive and negative control wells and the test sample color development well, and add another blank Add stop solution to the microplate plate, measure the OD value with a microplate reader, and judge the result. In the original reaction plate, add compound cleaning solution (0.5 mol / L H 2 SO 4 100ml+3ml ether), fill the reaction well and let it stand for 5 minutes to pour it out. Then wash with PBS buffer 3 times with an interval of 30 seconds each time, then it can be used to measure the next batch of samples. If you do not use this plate at that time, you can store it in a refrigerator at 4°C. The result will not change within a week. When you use it next time, first pat the plate dry, and then measure it according to the original method. Cycle more than two times, and then add th...

Embodiment 2

[0007] Example 2: Xiamen Xinchuang anti-HCV kit, batch number 971110, according to the operation manual, after adding the substrate for color development, add the stop solution, measure the OD value with a microplate reader to judge the result, then pour out the original reaction solution, add Composite cleaning solution (0.5 mol / liter HC1100ml mixed with 3ml ether and 3g urea) filled the reaction well, let it stand for 10 minutes and poured it off, repeated once, and then washed with PBS buffer 3 times, then it can be used to determine the next batch of samples, according to the original method The operation steps can be determined. If it is not used at that time, it can be placed in a refrigerator at 4°C for two weeks, and the results will not change. Add 1-2 wells of quality control material each time as a reference to determine the number of cycles.

Embodiment 3

[0008] Example 3: For the anti-HGV kit of Luoyang Huamei Biological Co., Ltd., according to the operation manual, add the stop solution after adding the substrate for color development, measure the OD value with a microplate reader, and judge the result. Afterwards, the original reaction solution was poured out, and the compound cleaning solution (0.5 mol / liter HCl100ml+3g urea) was added to fill the reaction well and left to stand for 5 minutes, then washed with PBS buffer three times to measure the next batch of samples. If it is not used at the right time, the result will not change within two weeks in a 4°C refrigerator. Add 1-2 wells of quality control substances for each measurement, as a reference to determine the number of cycles. The enzyme-substance conjugates of the above various methods should be pre-tested to determine the best potency after self-preparation, and the substrates can be prepared according to general routine methods.

[0009] The present invention is...

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Abstract

The present invention relates to a method and technique for regenerate the reaction board, tube or bead used for clincial diagnosis of hepatitis and AIDS viruses and bacteria, chlamydozoan and protozoon infection by enzyme-linked immunosobant assay (ELISA), which features use of a composite cleaning liquid prepared from sulfuric acid or hydrochloric acid or acetic acid and ether or urea to dissociate the antibody or antigen combined to the reaction board, so the reaction board can be reused. Its advantage is no influence to specificity and sensitivity.

Description

technical field [0001] The invention relates to a method for regenerating and reusing reaction plate tubes and beads used in the clinical diagnosis of hepatitis, AIDS and other viruses and other bacteria, chlamydia and protozoa infection by enzyme-linked immunosorbent assay (EL ISA method) and corresponding technology. Background technique [0002] Enzyme-linked immunosorbent assay (ELISA) is an important technology for clinical diagnosis and experimental research at home and abroad, involving many disciplines such as medicine, biology, genetics, and immunology. , with known antigens (antibodies) to detect unknown antibodies (antigens), it is widely used in the diagnosis of diseases and the observation of curative effects and preventive medicine, such as the diagnosis and differential diagnosis of hepatitis virus (A, B, C) in medicine , D, E, Hepatitis G virus), AIDS diagnosis and other bacteria, chlamydia, protozoa infection diagnosis. [0003] The reaction plates detected...

Claims

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Application Information

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IPC IPC(8): G01N33/53
Inventor 徐树良
Owner NANJING RED CROSS BLOOD CENT
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