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Method for eliminating HRP (Horseradish Peroxidase) antibody interference

A technology of capture method and enzyme-linked immunoassay, which is applied in the field of in vitro diagnosis and can solve problems such as interference with test results

Inactive Publication Date: 2019-12-31
AUTOBIO DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

When the antigen is tested by the competition method, when the sample to be tested contains HRP antibody, it will bind to the HRP-labeled antigen or HRP-labeled antibody, thereby interfering with the test result

Method used

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  • Method for eliminating HRP (Horseradish Peroxidase) antibody interference
  • Method for eliminating HRP (Horseradish Peroxidase) antibody interference
  • Method for eliminating HRP (Horseradish Peroxidase) antibody interference

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1. Effect of adding apo-HRP on TORCH-IgM detection results

[0024] 25 samples to be tested were detected by magnetic particle TORCH-IgM (TOX / RV / CMV / HSV-1 / HSV-2 IgM) 5-item detection kit (Antu Bio). In addition, use the magnetic particle TORCH-IgM kit (Antu Biology) with a blocking agent (0.5 mg / ml apo-inactivated HRP) added to the enzyme conjugate or sample diluent to simultaneously detect the above 25 samples to be tested. The results are shown in Table 1.

[0025] Table 1 compares the effect of adding apo-HRP on the detection results of TORCH-IgM

[0026]

[0027]

[0028] The results showed that after adding 0.5mg / ml apo-inactivated HRP, the false positives of TORCH-IgM multi-positive samples (containing HRP antibody in the samples) were eliminated, while the detection results of normal samples were not affected.

Embodiment 2

[0029] Example 2. Adding apo-HRP eliminates the interference of HRP antibody to the competition method

[0030] The progesterone quantitative detection kit (magnetic particle chemiluminescence method) of Antu Bio and the progesterone detection kit (electrochemiluminescence method) of Roche were used to detect 16 samples to be tested. In addition, the magnetic particle progesterone quantitative detection kit (Antu Biology) added with a blocking agent (0.01mg / ml apo-HRP) to the enzyme conjugate was used to simultaneously detect the above 16 samples to be tested. The test results are shown in Table 2 and Figure 1-2 .

[0031] Table 2 compares the effect of adding apo-HRP on the detection results of the competition method

[0032]

[0033]

[0034] The results showed that compared with the results detected by the progesterone quantitative detection kit (magnetic particle chemiluminescence method) without adding blocking agent, the amount detected by the progesterone quanti...

Embodiment 3

[0036] 1. Verification of the HRP antibody contained in the sample 1

[0037] The aprosthetic group-inactivated HRP was coated on the microtiter plate, after washing, 25 samples to be tested were added, after incubation and washing, enzyme-labeled anti-antibodies (anti-human IgG antibody, anti-human IgM antibody) were added for detection by indirect method IgG and IgM antibodies in the sample (IgM antibody detection: the sample is pre-adsorbed with goat anti-human IgG), the experimental data is as follows:

[0038] Table 3: Sample containing HRP antibody validation experiment 1

[0039]

[0040]

[0041]The results showed that there was no antibody against inactivated HRP in normal samples, but antibodies against inactivated HRP were contained in TORCH-IgM positive samples.

[0042] 2. Verification of the HRP antibody contained in the sample 2

[0043] Use the magnetic particle CMV-IgM antibody detection kit to detect 20 samples to be tested, add 0.01mg / ml HRP (with bi...

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Abstract

The invention belongs to the technology of in-vitro diagnosis, and discloses a method for eliminating HRP (Horseradish Peroxidase) antibody interference. A blocking agent is added into an immunodetection system taking horseradish peroxidase as a marker, and the blocking agent is inactivated HRP with a prosthetic group being removed. The combination of the inactivated HRP with the prosthetic groupbeing removed and the HRP antibody can block the combination of the HRP antibody and an HRP labeled antibody or an HRP labeled antigen in the serum, so that the interference of the HRP antibody on thedetection result is eliminated. According to the method for eliminating HRP antibody interference, the common problem existing in the immunodetection system taking horseradish peroxidase as a markeris solved, false positive caused by HRP antibody interference is eliminated, especially multi-item IgM false positive occurring during IgM joint detection is eliminated, and the problem of poor magnitude correlation caused by HRP antibody interference in immunoassay can also be improved. The addition of the blocking agent does not influence the detection result of a normal sample and improves theaccuracy of the detection result.

Description

technical field [0001] The invention belongs to in vitro diagnostic technology, in particular to a method for eliminating HRP antibody interference, in particular to a method for eliminating HRP antibody interference in an immunoassay system using horseradish peroxidase as a marker. Background technique [0002] Horseradish Peroxidase (HRP) is a glycoprotein with a molecular weight of 44,000 and containing heme. It is widely distributed in the plant kingdom, especially in horseradish, because it is also called horseradish. peroxidase. As early as the 1930s, someone started to isolate this enzyme from horseradish, and later prepared crystals. Usually, horseradish is used as raw material in the test, after water extraction, ammonium sulfate and acetone fractionation, then zinc ion purification, dialysis for desalination, and freeze-drying, high-purity horseradish peroxidase can be obtained. Markers are the key reagents in immunoenzyme technology. Due to the advantages of eas...

Claims

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Application Information

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IPC IPC(8): G01N33/535G01N33/543G01N33/64
CPCG01N33/535G01N33/54326G01N33/64
Inventor 王万利王春霞陶占领刘功成张跃峰渠海郑业焕付光宇吴学炜
Owner AUTOBIO DIAGNOSTICS CO LTD
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