Chemical luminescence method immune analysis diagnostic reagent kit for detecting cytomegalovirus IgG 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

AI Technical Summary

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

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Preparation of a chemiluminescent immunoassay diagnostic kit for detecting cytomegalovirus IgG antibody:

[0048] A. Coated plate preparation:

[0049] Coat the microwell plate with 50mmol / L pH9.6 carbonate buffer solution containing 2.5μg / ml FITC antibody at a volume of 100μL / well. After coating for 18-20 hours at 2~8℃, use (weight) 20mmol / L pH7.4 Tris-HCl buffer solution 180μL / well to seal for 18-20 hours, shake off the blocking solution, dehumidify, dry at 28°C for 18-20 hours, add desiccant to seal, and place in 2-8 Store at ℃.

[0050] B. Preparation of FITC-labeled cytomegalovirus antigen:

[0051] a) FITC labeling method: dialyze 1mg of cytomegalovirus antigen with 50mmol / L pH9.6 carbonate buffer at 2-8°C overnight, and mix 1mg / mL with the ratio of cytomegalovirus antigen: FITC=1mg:200μg Slowly add FITC (dimethyl sulfoxide as solvent) solution into the cytomegalovirus antigen solution and mix evenly, and react for 4 hours at 2-8°C in the dark, then add 5mol / L ...

Embodiment 2

[0071] Preparation of a chemiluminescent immunoassay diagnostic kit for detecting cytomegalovirus IgG antibody:

[0072] Using magnetic particles as a solid phase carrier, the FITC antibody was covalently coupled to the surface of the magnetic particles by the EDC method, and the 50mmol / L pH7.2 Tris-HCl buffer containing 12% (weight) NaCl and 0.1% (weight) KCl was used as the Concentrate the washing solution, and prepare the chemiluminescent immunoassay diagnostic kit for cytomegalovirus IgG antibody of this example in the same manner as in Example 1 for the rest.

Embodiment 3

[0074] Preparation of a chemiluminescent immunoassay diagnostic kit for detecting cytomegalovirus IgG antibody:

[0075] (1) Preparation of luminescent substrate solution:

[0076] Dissolve 100ml of AMPPD, 120g of NaCl, 1g of KCl, and 6.05g of tris (Tris) in 900ml of double-distilled water, adjust the pH to 8.0-10.0 with HCl, and adjust the solution to 1000mL with double-distilled water.

[0077] (2), the preparation of concentrated washing liquid:

[0078] Dissolve 120g NaCl, 1g KCl, 6.05g Tris, 0.5ml Tween 20 in 900ml double-distilled water, adjust the pH to 7.2 with HCl, and dilute to 1000mL with double-distilled water.

[0079] Dilute 20 times with double distilled water.

[0080] All the other preparation methods are the same as in Example 1.

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Abstract

The invention discloses a chemiluminescence immunoassay analysis diagnosing test kit and a preparing method thereof for detecting cytomegalovirus IgG antibody. The test kit includes positive and negative reference substance, solid-phase carrier, labeled antigen, enzyme-labeled antibody, chemiluminescence zymolyte liquid and condensed washing liquid. The test kit has great significance in screening diagnosis and monitoring for pre-pregnancy and early-pregnancy infection of childbearing women as well as CMV infection diagnosis of high risk group (such as tumour, organ transplanting, HIV and the like ).

Description

technical field [0001] The invention relates to the field of medical immunological in vitro diagnostic reagents, and more specifically relates to a diagnostic kit for detecting cytomegalovirus IgG antibody and a preparation method thereof by combining FITC antibody-FITC indirect coating technology with chemiluminescence immunoassay. Background technique [0002] Cytomegalovirus (Cytomegalovirus.CMV) is a virus that Smith isolated from the submandibular gland of a dead baby in 1956. It was officially named cytomegalovirus in 1964. It belongs to the herpesviridae family and is a double-stranded DNA virus. A widespread species-specific virus. CMV infection is distributed worldwide, and humans are the only host of CMV, and its prevalence varies with regions and socioeconomic conditions. The positive rate of serum CMV antibody in the adult population of North America and Europe is 50-80%, while the positive rate of serum antibody in the population of Asia and Africa is nearly 10...

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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