Homogeneous immunodetection kit for detecting target immunoglobulin M (IgM) antibody in sample as well as using method and application of homogeneous immunodetection kit

A technology of samples and kits, which is applied in the field of biomedical testing, can solve problems such as false positives, reduce work efficiency, and take a long time, and achieve the effect of solving influence and removing interference

Active Publication Date: 2018-05-01
CHEMCLIN DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method has the following two disadvantages: one is that the anti-human IgM antibody is used as a capture antibody and cannot distinguish between specific IgM antibodies and non-specific IgM antibodies. When the concentration of specific IgM antibodies is low and the concentration of non-specific IgM antibodies is particularly high, it may Presence of specific IgM antibodies is removed by washing, resulting in false negative results
The second is that the detection process requires two steps of incubation and washing. In the washing process, it is very dependent on the washing efficiency of the washing ...

Method used

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  • Homogeneous immunodetection kit for detecting target immunoglobulin M (IgM) antibody in sample as well as using method and application of homogeneous immunodetection kit
  • Homogeneous immunodetection kit for detecting target immunoglobulin M (IgM) antibody in sample as well as using method and application of homogeneous immunodetection kit
  • Homogeneous immunodetection kit for detecting target immunoglobulin M (IgM) antibody in sample as well as using method and application of homogeneous immunodetection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] Embodiment 1: the detection of HBc IgM antibody

[0079] The reagents used include:

[0080] R1: a solution of complement C1q connected to the receptor microspheres at a concentration of 25ug / ml;

[0081] R2: a solution of HBcAg linked to biotin with a concentration of 1 ug / ml;

[0082] R3: diluent containing IgG antibody blocking agent;

[0083] R4: a solution of donor microspheres linked to streptavidin (SA) at a concentration of 25 ug / ml.

[0084] The preparation method, composition structure and content of the acceptor microspheres used in the present invention can be referred to Example 1 of Chinese patent CN100429197C, and then the C1q complement is coated on the surface of the acceptor microspheres.

[0085] Donor microspheres are to put 200g chlorophyll A into 200nm carboxy-modified latex particles according to the method described in the examples of patent US5780646, and coat the surface with streptavidin to form the donor microspheres described in the prese...

Embodiment 2

[0093] Embodiment 2: the detection of HEV IgM antibody

[0094] The reagents used include:

[0095] R1: a solution of HEV Ag connected to acceptor microspheres with a concentration of 25ug / ml;

[0096] R2: a biotin-linked complement C1q solution with a concentration of 1 ug / ml;

[0097] R3: diluent containing IgG antibody blocking agent;

[0098] R4: Streptavidin-linked donor microsphere solution at a concentration of 25 ug / ml.

[0099] The detection steps are:

[0100] (1) Add 100ul of reagent R3 to the reaction well, then add 10ul of the sample to be tested, and incubate at 37°C for 5min;

[0101] (2) Add 25ul each of reagent R1 and reagent R2 to the reaction well, and incubate at 37°C for 10min to obtain the first mixture;

[0102] (3) Add 25ul of reagent R4 to the reaction well and incubate at 37°C for 10min to obtain the second mixture;

[0103] (4) Put the second mixture into the light-excited chemiluminescence detection instrument, and read the chemiluminescence s...

Embodiment 3

[0106] Embodiment 3: the detection of HBc IgM antibody

[0107] The reagents used include:

[0108] R1: a solution of complement C1q connected to the receptor with a concentration of 25ug / ml;

[0109] R2: a solution of HBcAg linked to biotin with a concentration of 1 ug / ml;

[0110] R3: diluent containing IgG antibody blocking agent;

[0111] R4: Donor solution linked to streptavidin (SA) at a concentration of 25 ug / ml.

[0112] The complement C1q used in the present invention connected to the receptor is prepared according to the scheme described in the patent PCT / US2010 / 025433, and its structure is composed of C1q-BSA-(dimethylthiophene) (BHHCT), in aqueous solution can be completely dissolved in.

[0113] The donor is to put 200g of chlorophyll A into 200nm carboxy-modified latex particles according to the method described in the examples of the patent US5780646, and coat the surface with streptavidin to form the donor microparticles of the present invention. ball.

...

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Abstract

The invention relates to a homogeneous immunodetection kit for detecting a target immunoglobulin M (IgM) antibody in a sample as well as a using method and application of the homogeneous immunodetection kit. The kit comprises a reagent a, a reagent b and a reagent c, wherein the reagent a comprises a known antigen, and the known antigen can be specifically bonded to the target IgM antibody in thesample to be detected so as to form a first complex; the reagent b comprises a complement C1q, the complement C1q can be specifically bonded to the first complex so as to form a second complex, and the complement C1q is not enabled to be bonded to a free IgM antibody; any of the known antigen and the complement C1q is connected with a receptor; the receptor can react with received singlet oxygen so as to produce detectable chemiluminescence signals; the reagent c comprises a donor, and the donor can generate the singlet oxygen in an excited state. A method for detecting the target IgM antibodyin the sample by using the kit overcomes the effect of a non-specific IgM antibody on detection.

Description

technical field [0001] The invention belongs to the technical field of biomedical detection, and in particular relates to a homogeneous immunoassay kit for detecting target IgM antibodies in a sample and its use method and application. Background technique [0002] The detection of IgM antibodies is of great significance in medical testing. When a pathogen first infects the body, the first antibody produced is always IgM antibody. With the progress of the body's immune response, antibody class switching occurs to produce IgG, IgM or IgA antibody. Therefore, in the diagnosis of infectious diseases, a positive IgM antibody test is a sign of current infection. [0003] At present, the commonly used IgM antibody detection method is the capture method, which is to coat the anti-human IgM (μ chain) antibody on the solid phase carrier, and after the serum / plasma sample reacts with the solid phase carrier, all the IgM antibody in the sample Captured on the solid phase carrier, aft...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N33/68
CPCG01N33/56983
Inventor 李会强赵文雅刘宇卉李临
Owner CHEMCLIN DIAGNOSTICS CO LTD
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