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Kit for detecting an EB viral capsid antigen IgA antibody

An EB virus and antibody detection technology, applied in biological tests, measuring devices, material inspection products, etc., can solve the problems of low degree of automation, high subjectivity, and large differences in result reporting, and achieve automatic detection, avoiding specificity and sensitivity. Affected effect

Inactive Publication Date: 2019-04-16
AUTOBIO DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Immunoenzyme method requires immobilized cells as antigens, and there are large differences between batches. Microscopic observation results are highly subjective in judgment, and the results reported by different institutions vary greatly. It is difficult to perform quality control and parallel comparison. In addition, specialized technical personnel are required to operate. prone to errors
Enzyme-linked immunoassay is to immobilize antigens or antibodies on polystyrene microwell plates, use horseradish peroxidase or alkaline phosphatase as markers, and judge the results by color absorbance value. The reaction time of the experiment is long and it is automated. low level
Colloidal gold chromatography immobilizes antigens or antibodies on nitrocellulose membranes, uses colloidal gold as a marker, and develops color on the detection line through liquid capillary action. The interpretation of the results basically requires visual observation, which is fast but has low accuracy.

Method used

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  • Kit for detecting an EB viral capsid antigen IgA antibody
  • Kit for detecting an EB viral capsid antigen IgA antibody
  • Kit for detecting an EB viral capsid antigen IgA antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 Preparation of Epstein-Barr virus capsid antigen IgA antibody detection kit

[0022] 1. Preparation of VCA antigen-coupled magnetic particle suspension

[0023] 1.1 Magnetic particle washing

[0024] Take 200ul of magnetic particles containing carboxyl groups on the surface and put them in a glass bottle, use a magnet to adsorb the magnetic particles to the bottom of the glass bottle, remove the supernatant; add 2ml of 0.02M PBS (pH 8.0), repeat the above operation 3 times.

[0025] 1.2 Activation of magnetic particles

[0026] Dissolve EDC and NHS in 0.1M MES (pH 5.0) buffer solution at a concentration of 10-70mg / ml, then add 1ml of each to the magnetic particles; shake gently at room temperature for 30-60 minutes; The particles are adsorbed at the bottom, remove the supernatant; then add 2ml 0.1M MES (pH 5.0) buffer to resuspend the magnetic particles; repeat the above operation twice.

[0027] 2. Preparation of anti-human IgA monoclonal antibody enzyme c...

Embodiment 2

[0036] Embodiment 2 The detection method of kit of the present invention

[0037] The kit prepared in Example 1 was used in conjunction with the automatic chemiluminescence instrument AutoLumo A2000 or AutoLumo A2000 Plus produced by Zhengzhou Antu Bioengineering Co., Ltd. to detect respiratory syncytial virus IgM antibody:

[0038] Add 3 wells of positive control (for determining the Cutoff value) and 2 wells of negative control in sequence in the reaction container (hereinafter referred to as "well"), 100 µL / well; add 10 µL sample and 100 µL sample diluent to each well of the remaining wells;

[0039]Add 20 µL of magnetic particle suspension to each well; mix well, incubate at 37°C for 15 minutes; then wash with washing solution 5 times;

[0040] Add 50 µL of enzyme conjugate and 50 µL of antigen solution to each well; mix well and incubate at 37°C for 17 minutes; wash with cleaning solution 5 times;

[0041] Finally, add 50 µL each of Substrate A and Substrate B to each we...

Embodiment 3

[0045] Embodiment 3 Performance evaluation of the kit of the present invention

[0046] 1. Repeatability

[0047] Repeatability refers to the repeated measurement of the same sample, and the closeness of each measurement result to the mean value, expressed in the coefficient of variation CV.

[0048] Select 3 samples with different concentrations, test 4 repetitions every day, and continuously test for 5 days, the coefficient of variation CV<8%, the results are shown in Table 1:

[0049] Table 1

[0050]

[0051] From the results in Table 1, it can be seen that the coefficient of variation of sample 1 is 3.95%, the coefficient of variation of sample 2 is 4.25%, and the coefficient of variation of sample 3 is 6.72%, all of which are less than 10% required by the industry, which proves that its repeatability is good.

[0052] 2. Specificity

[0053] Detect hepatitis B, hepatitis C, syphilis, HIV, EB-EA, EB-NA1, HSV-1, HSV-2, CMV, VCA-IgG, VCA-IgM and HAMA positive samples,...

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Abstract

The invention discloses a kit for detecting an EB viral capsid antigen (VCA) IgA antibody. The kit comprises a magnetic particle suspension taking a VCA gene recombinant antigen fragment as a coatingantigen, an anti-human IgA monoclonal antibody enzyme conjugate marked by horse radish peroxidase, a sample diluent and a chemiluminescent substrate solution. Magnetic particles are adopted as a solid-phase carrier of the antibody, and a chemiluminescence immunoassay technology is utilized, so that single-person random, rapid and automatic detection can be realized on a full-automatic chemiluminescence analyzer. Due to the fact that an indirect method is adopted in the technical principle, a plurality of dominant antigen fragments most valuable to previous infection and recent infection are selected from more than 30 EB virus target antigen proteins, and the problem that specificity and sensitivity are affected due to incomplete sites of conventional kits on the market is solved. The kit is suitable for the qualitative detection of the EB VCA IgA antibody in a human serum or plasma sample, and assists the clinical diagnosis of nasopharyngeal carcinoma.

Description

technical field [0001] The invention relates to in vitro diagnostic technology, in particular to an Epstein-Barr virus capsid antigen (VCA) IgA antibody detection kit. Background technique [0002] Epstein-Barr virus is the only lymphoid follicular virus that can cause human infection in the gamma subfamily of the Herpesviridae virus family. It has the characteristics of B lymphocytes and can establish a recessive infection in B lymphocytes and stimulate cell proliferation and transformation. It may be associated with a variety of diseases including infectious mononucleosis, Burkitt's lymphoma, Burkits' sarcoma, Hodgkin's disease, and nasopharyngeal carcinoma, among others. Among them, 80% of nasopharyngeal carcinoma cases occur in China, and the morbidity and mortality of nasopharyngeal carcinoma rank first among malignant tumors in several provinces in southern my country. Capsid antigen (VCA) IgA antibody is produced during the cleavage and replication phase of cells inf...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/577G01N33/536G01N33/543G01N21/76
CPCG01N21/76G01N33/536G01N33/54326G01N33/577G01N33/6893
Inventor 孙冯博王新明刘功成李晓霞乔晓芳郑业焕渠海付光宇吴学炜
Owner AUTOBIO DIAGNOSTICS CO LTD
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