阅读说明：本技术 一种消除hrp抗体干扰的方法 (Method for eliminating interference of HRP antibody ) 是由 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 于 2019-09-20 设计创作，主要内容包括：本发明属于体外诊断技术,公开了一种消除HRP抗体干扰的方法,在以辣根过氧化物酶为标记物的免疫检测系统中添加阻断剂,所述阻断剂为脱辅基失活的HRP。脱辅基失活的HRP与HRP抗体结合能阻断血清中的HRP抗体与HRP标记抗体或HRP标记抗原的结合,从而消除HRP抗体对检测结果的干扰。本发明所述消除HRP抗体干扰的方法,解决了以辣根过氧化物酶为标记物的免疫检测系统中存在的共性问题,消除HRP抗体干扰导致的假阳性,特别是消除IgM联检时出现的多项目IgM假阳性,也能改善免疫分析中由于HRP抗体干扰导致量值相关性差问题。且添加阻断剂不影响正常样本的检测结果,提升检测结果的准确性。(The invention belongs to an in-vitro diagnosis technology and discloses a method for eliminating HRP antibody interference. The combination of the inactivated apoprotein HRP and the HRP antibody can block the combination of the HRP antibody in serum and the HRP labeled antibody or the HRP labeled antigen, thereby eliminating the interference of the HRP antibody on the detection result. The method for eliminating the interference of the HRP antibody solves the common problem in an immunodetection system taking horseradish peroxidase as a marker, eliminates false positive caused by the interference of the HRP antibody, especially eliminates multi-project IgM false positive generated in IgM joint detection, and can also improve the problem of poor value correlation caused by the interference of the HRP antibody in immunoassay. And the addition of the blocking agent does not affect the detection result of a normal sample, so that the accuracy of the detection result is improved.)

1. A method for eliminating interference of HRP antibody is characterized in that a blocking agent is added into an immunoassay system taking horseradish peroxidase as a marker, and the blocking agent is apoplectic inactivated HRP.

2. The method of claim 1, wherein the apo-inactivated HRP is added at a concentration of 0.01-0.5 mg/ml.

3. The method of claim 1, wherein the apo-inactivated HRP additive component is in a sample diluent or an enzyme conjugate.

4. The method of claim 1, wherein the immunoassay system using horseradish peroxidase as a label comprises an enzyme-linked immunoassay system using horseradish peroxidase as a label and a chemiluminescent assay system using horseradish peroxidase as a label.

5. The method according to claim 4, wherein the detection method of the enzyme-linked immunosorbent assay or chemiluminescence assay detection system is IgM detection by a capture method or antigen detection by a competition method.

6. The method of claim 5, wherein the IgM determined by the capture method is IgM determined by an enzyme-labeled antigen method or IgM determined by an enzyme-labeled antibody plus antigen method.

Technical Field

The invention belongs to an in-vitro diagnosis technology, and particularly relates 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

Horseradish Peroxidase (HRP) is a glycoprotein with a molecular weight of 44000 and containing heme, is widely distributed in the plant world, and particularly has high content in Horseradish, and is also called Horseradish Peroxidase. The isolation of this enzyme from horseradish was undertaken as early as the 30 s in the 20 th century, after which crystals were prepared. In general, horseradish is used as a raw material in the test, and the horseradish peroxidase with high purity can be obtained by water extraction, ammonium sulfate and acetone fractional separation, zinc ion purification, dialysis and desalination and freeze drying. The marker is a key reagent in the immunoenzyme technology, and the HRP becomes the most widely applied marker in the ELISA experiment due to the advantages of easy extraction, low price, stable property and the like.

Horseradish is a cold-resistant perennial herb, mainly distributed in temperate regions of the world, and the root of the herb has good cooking value. People have a long history of eating horseradish, and the traditional dishes of many nationalities show the shadow of the horseradish. As early as about 1 century, the horse radish has appeared in the roman menu, and in about 13 th century, the horse radish is introduced into Germany and gradually becomes the most important flavoring in Europe, and the kosher traditional food "kosher horse radish sauce" is prepared by mixing horse radish mud with vinegar, sugar and salt. The horse radish paste is mainly prepared by crushing horse radish into powder and adding water, is a seasoning for dipping uncooked seafood and horse meat, and is widely applied in Japan. Horseradish is introduced into Shanghai by English people 80 years ago, at present, the Shanghai, Jiangsu, Shandong, Liaoning and the like are cultivated in China, and most of horseradish sauce and horseradish powder sold in China at present are made of the horseradish. Since horseradish is a food material, antibodies against horseradish, i.e., HRP, may be present in the human body.

When IgM is detected by the capture method, when IgM antibodies resisting HRP are contained in a sample to be detected, the IgM antibodies are captured on a coating plate or magnetic particles, and the IgM antibodies are combined with HRP-labeled antigen or HRP-labeled antibody to generate false positive. In the competitive method for detecting antigen, when the sample to be detected contains HRP antibody, the HRP antibody is combined with the HRP-labeled antigen or the HRP-labeled antibody, so that the detection result is interfered.

At present, domestic manufacturers basically adopt a capture method for detecting the optimal birth and care TORCH IgM antibodies, and clinically, the TORCH-IgM 5 antibodies are all positive. The TORCH infection screening is the infection screening of a series of pathogens, and the pathogens have no infection correlation, so that the phenomenon that one sample is simultaneously and acutely infected with a plurality of viruses is very rare clinically, the probability that IgM (immunoglobulin M) homoyang is generally false positive is very high, and IgM (immunoglobulin M) homoyang samples are basically false positive after being confirmed. Therefore, a method for eliminating interference of HRP antibodies is needed to be found, which is used for eliminating false positives caused by interference of HRP antibodies in an immunoassay system using HRP as a marker.

Disclosure of Invention

In view of this, the present invention provides a method for eliminating HRP antibody interference, which is used to eliminate false positives caused by HRP antibody interference in an immunoassay system using horseradish peroxidase as a marker, and to improve the accuracy of detection results.

In order to achieve the purpose, the invention can adopt the following technical scheme:

a method for eliminating interference of HRP antibody is characterized in that a blocking agent is added into an immunoassay system taking horseradish peroxidase as a marker, and the blocking agent is apoplectic inactivated HRP.

Apocynated HRP is a product of HRP removal of prosthetic heme, which is biologically inactive with HRP. According to the method for eliminating the interference of the HRP antibody, the apoplectic inactivated HRP is added into an immunodetection system taking horseradish peroxidase as a marker, and the binding of the apoplectic inactivated HRP and the HRP antibody can block the binding of the HRP antibody in serum and the HRP labeled antibody or HRP labeled antigen, so that the interference of the HRP antibody on a detection result is eliminated.

Preferably, the apo-inactivated HRP is added in a concentration of 0.01-0.5mg/ml in the method for eliminating interference of HRP antibody.

In some embodiments, the apo-inactivated HRP is added at a concentration of 0.5mg/ml in the method of eliminating HRP antibody interference.

In the method for eliminating HRP antibody interference, the added components of the apoplectic HRP are sample diluent or enzyme conjugate. That is, the apoprotein-inactivated HRP is added to a sample diluent or an enzyme conjugate in an immunoassay system using horseradish peroxidase as a marker.

As will be understood by those skilled in the art, the immunoassay system using horseradish peroxidase as a label includes an enzyme-linked immunoassay system using horseradish peroxidase as a label and a chemiluminescent assay system using horseradish peroxidase as a label.

The detection method of the enzyme-linked immunosorbent assay or chemiluminescence assay detection system comprises but is not limited to a capture method for IgM detection or a competition method for antigen detection.

The capture method for measuring IgM includes, but is not limited to, measuring IgM by an enzyme-labeled antigen method or measuring IgM by an enzyme-labeled antibody plus antigen method.

The invention discloses a method for eliminating HRP antibody interference, which is characterized in that a blocking agent is added into an immunodetection system taking horseradish peroxidase as a marker, and the blocking agent is an apoplectic inactivated HRP. The combination of the inactivated apoprotein HRP and the HRP antibody can block the combination of the HRP antibody in serum and the HRP labeled antibody or the HRP labeled antigen, thereby eliminating the interference of the HRP antibody on the detection result. The method for eliminating HRP antibody interference relates to IgM antibody detection by a capture method, antigen detection by a competition method and the like, solves the common problem existing in an immunodetection system using horseradish peroxidase as a marker, eliminates false positive caused by HRP antibody interference, especially eliminates multinomial IgM false positive generated in IgM joint detection, and can also improve the problem of poor value correlation caused by HRP antibody interference in immunoassay. And the addition of the blocking agent does not affect the detection result of a normal sample, so that the accuracy of the detection result is improved.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below.

FIG. 1 is a graph showing the correlation between the measurement values of the quantitative determination kit for progesterone without a blocking agent (magnetic particle chemiluminescence method) and the measurement values of the progesterone detection kit for Roche (electrochemiluminescence method) in EXAMPLE 2;

FIG. 2 is a graph showing the correlation between the measurement values of the blocking agent-added progesterone quantitative assay kit (magnetic particle chemiluminescence method) and Roche progesterone assay kit (electrochemiluminescence method) in EXAMPLE 2.

Detailed Description

The invention discloses a method for eliminating interference of an HRP antibody. Those skilled in the art can modify the process parameters appropriately to achieve the desired results with reference to the disclosure herein. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the methods and products of the present invention have been described in terms of preferred embodiments, it will be apparent to those of ordinary skill in the art that variations and modifications of the methods described herein, as well as other suitable variations and combinations, may be made to implement and use the techniques of the present invention without departing from the spirit and scope of the invention.

In order to further understand the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Unless otherwise specified, the reagents involved in the examples of the present invention are all commercially available products, and all of them are commercially available.