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Chemical luminescence method immune analysis diagnostic reagent kit for detecting cytomegalovirus IgM antibody

A chemiluminescence immunity, cytomegalovirus technology, applied in chemiluminescence/bioluminescence, analysis by chemical reaction of materials, analysis of materials, etc., can solve the problems of toxic substrates, low sensitivity, inconvenient operation, etc. Simple operation, high sensitivity and high compliance

Inactive Publication Date: 2009-02-18
CHEMCLIN DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0011] In order to solve the problems of inconvenient operation, low sensitivity, short validity period of radioimmunoassay reagents, pollution hazards of radioactive waste, interference of enzyme immunoassay reagents by various factors, and most toxic substrates in clinical etiology tests, the present invention provides a method using FITC antibody-FITC indirect coating technology combined with chemiluminescent immunoassay technology to detect cytomegalovirus IgM antibody in vitro diagnostic kit and preparation method thereof

Method used

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  • Chemical luminescence method immune analysis diagnostic reagent kit for detecting cytomegalovirus IgM antibody

Examples

Experimental program
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Effect test

Embodiment 1

[0060] Preparation of cytomegalovirus IgM antibody chemiluminescence immunoassay diagnostic kit:

[0061] A. Preparation of coated plate:

[0062] Coat the microplate with 50 mmol / L pH 9.6 carbonate buffer solution containing 2 μg / ml FITC antibody at a volume of 100 μL / well. 180 μL / well of 20 mmol / L pH7.4 Tris-HCl buffer solution of 180 μL / well was blocked for 18 to 20 hours, drained off the blocking solution, dehumidified, dried at 28 °C for 18 to 20 hours, sealed with desiccant, and stored at 2 to 8 °C.

[0063] B. Preparation of FITC-labeled antibody:

[0064] a) FITC labeling: 1 mg of anti-human IgM monoclonal antibody was dialyzed against 50 mmol / L pH 9.6 carbonate buffer at 2-8 °C, and FITC was dissolved in dimethyl sulfoxide (DMSO) at a concentration of 1 mg / mL , according to the ratio of cytomegalovirus antigen:FITC=1mg:150μg, slowly add FITC dimethyl sulfoxide solution to cytomegalovirus antigen solution and mix well, react at 4°C for 8 hours in the dark, add 5mol / L...

Embodiment 2

[0084] Preparation of cytomegalovirus IgM antibody chemiluminescence immunoassay diagnostic kit:

[0085] Using magnetic particles as solid phase carrier, the FITC antibody was covalently coupled to the surface of magnetic particles by glutaraldehyde method. In order to concentrate the washing solution, the rest were prepared in the same manner as in Example 1 to prepare a cytomegalovirus IgM antibody chemiluminescence immunoassay diagnostic kit.

Embodiment 3

[0087] Preparation of cytomegalovirus IgM antibody chemiluminescence immunoassay diagnostic kit:

[0088] (1) Preparation of luminescent substrate solution:

[0089] Dissolve 100ml AMPPD, 120g NaCl, 1g KCl, 6.05g tris (Tris) in 900ml double-distilled water, adjust the pH to 8.0-10.0 with HCl, and make up to 1000mL with double-distilled water.

[0090] (2), the preparation of concentrated washing solution:

[0091] Dissolve 120g NaCl, 1g KCl, 6.05g Tris (Tris) and 0.5ml Tween 20 in 900ml double distilled water, adjust the pH to 7.2 with HCl, and make up to 1000mL with double distilled water.

[0092] Dilute 20 times with double distilled water when using.

[0093] The remaining steps are the same as in Example 1.

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Abstract

The invention discloses an in-vitro diagnostic test kit and a preparing method thereof for detecting cytomegalovirus IgM antibody by adopting the FITC antibody -FITC indirectly coating technique combined with the chemiluminescence immunoassay technique. The test kit comprises positive and negative reference substance, solid-phase carrier, labeled antibody, enzyme-labeled antigen, chemiluminescence zymolyte liquid and condensed washing liquid. The test kit can be taken as the assistant detection critetia in prenatal diagnosis for eugenics, and has great significance in enhancing the birth population quality, doing well in family planning and eugenics.

Description

technical field [0001] The present invention relates to the field of medical immune in vitro diagnostic reagents, more particularly, to a diagnostic kit for detecting cytomegalovirus IgM antibody using FITC antibody-FITC indirect coating technology combined with chemiluminescence immunoassay and a preparation method thereof. Background technique [0002] Cytomegalovirus (Cytomegalovirus.CMV) is a virus isolated by Smith from the submandibular gland of a dead baby in 1956. It was officially named Cytomegalovirus in 1964. It belongs to the family Herpesviridae and is a double-stranded DNA virus. A widespread species-specific virus. CMV infection has a worldwide distribution, with humans being the sole reservoir of CMV, and its prevalence varies by region and socioeconomic status. Serum CMV antibody positivity rates in North American and European adult populations range from 50 to 80%, whereas seroprevalence in Asian and African populations is nearly 100%. [0003] Adult CMV ...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/569G01N21/76G01N33/535
Inventor 彭京胜应希堂宋胜利胡国茂唐宝军于尚永
Owner CHEMCLIN DIAGNOSTICS CO LTD
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