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Diagnosis reagent for syphilis spirochete antibody

A technology of Treponema pallidum and diagnostic reagents, which is applied in the field of Treponema pallidum antibody enzyme-labeled diagnostic reagents, can solve problems such as clinical judgment errors, burdens, and increased workload, and achieve standardized management assurance, good stability, and stable reagents.

Inactive Publication Date: 2003-03-26
肖洪武
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the instability of its chromogenic system, the chromogenic substrates TMB and hydrogen peroxide are usually stored separately and used dropwise, which doubles the workload of large-scale blood source screening, and is even more dangerous in densely packed enzyme labels. If one of the chromogenic substrates is omitted in the microwell, the test result will be false negative. If the syphilis positive serum is false negative, the harm caused by blood transfusion infection can be imagined.
[0005] TMB is the commonly used chromogenic agent of ELISA reagent, because 3,3 ', 5,5'-tetramethylbenzidine (abbreviated TMB) is insoluble in water, usually need to add a large amount of solvents to increase its solubility; and in the reaction system Another chromogenic substrate, hydrogen peroxide, is easy to turn blue when mixed together, causing clinical judgment errors (beyond the CutOff value), causing burden and waste to patients and hospitals

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0011] Reagent composition:

[0012] 1. Use synthetic peptide antigen or antigen expressed by genetic engineering: P15, P17, P47 antigen 1μg / ml coated on the microplate.

[0013] 2. Enzyme-labeled secondary antibody (IgG, IgM): containing 1:35K enzyme-labeled secondary antibody dilution solution

[0014] 3. Two-in-one color developer:

[0015] 0.05 parts of TMB sodium propanesulfonate;

[0016] 0.05 part of cumene hydroperoxide;

[0017] 10 parts of N-methyl-2-pyrrolidone;

[0018] 10 parts of ethanol;

[0019] 0.05 parts of ProClin300 (2-methyl-4-isothiazolin-3-one);

[0020] Piperazine-N, N'-bis(2-ethanesulfonic acid) buffer solution 20mmol / L pH7.0.

Embodiment 2

[0022] Reagent composition:

[0023] 1. Coating with synthetic peptide antigen or antigen expressed by genetic engineering: P15, P17, P47 antigen 1 g / ml

[0024] on a microtiter plate.

[0025] 2. Enzyme-labeled secondary antibody (IgG, IgM): containing 1:35K enzyme-labeled secondary antibody dilution solution

[0026] 3. Two-in-one color developer:

[0027] 0.05 parts of TMB sodium propanesulfonate;

[0028] Hydroxyurea 0.05 part;

[0029] Hydroxypropyl cyclodextrin-5mmol / L;

[0030] 10 parts of dimethyl sulfoxide;

[0031] 0.05 parts of ProClin300 (2-methyl-4-isothiazolin-3-one);

[0032] Piperazine-N, N'-bis(2-ethanesulfonic acid) buffer solution 20mmol / L pH7.0.

Embodiment 3

[0057] Reagent composition:

[0058] 1. Use synthetic peptide antigen or antigen expressed by genetic engineering: P15, P17, P47 antigen 1μg / ml coated on the microplate.

[0059] 2. Enzyme-labeled secondary antibody (IgG, IgM): containing 1:35K enzyme-labeled secondary antibody dilution solution

[0060] 3. Two-in-one color developer:

[0061] 0.5 parts of TMB sodium propanesulfonate;

[0062] Hydroxyurea 0.01 part;

[0063] 50% parts of N-methyl-2-pyrrolidone;

[0064] 0.5 parts of ProClin300 (2-methyl-4-isothiazolin-3-one);

[0065] Piperazine-N, N'-bis(2-ethanesulfonic acid) buffer solution 100mmol / L pH8.0.

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PUM

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Abstract

This invention discloses a diagnostic reagent of syphilis spirochete antibody enzyme labeling for diagnosing syphilis spirochete infection, containing synthetic peptide antigen or genetic engineering expressed antigen: antigen P15, P17, P47, 0.1-10 mg / ml coated on the enzyme labeling, the enzyme labeling two antigens (IgG, IgM): containing 1:20-1:100K their diluents solution and color-developing agent combination and solubilizer in which the said color-developing agent in the combination is TMB. The said combination can include combination of TMB with hydrogen peroxide cumon ir its derivatives or TMB with hydroxyurea. On the bases of applied synthetic peptide antigen and genetic engineering expressing antigen, a TMB saline is used in the diagnostic reagent with excellent water solubility and stable performance, matched with proper solubilizers, stabilizers to get much better stable performance.

Description

technical field [0001] The invention relates to a Treponema pallidum antibody enzyme-labeled diagnostic reagent for diagnosing Treponema pallidum infection. technical background [0002] Syphilis is a complex sexually transmitted disease caused by Treponema pallidum (Spirillum). The route is vertical transmission through the blood. The course of syphilis usually includes primary, secondary and latent periods. Symptoms of tertiary syphilis can occur 1-30 years after primary infection and include gumma lesions, cardiovascular syphilis, and neurologic syphilis. Since Treponema pallidum cannot be cultured in vitro, the diagnosis can only rely on clinical signs, direct observation of bacteria at the injury site, and serological methods. Serological methods are suitable for the diagnosis of all stages of syphilis except very early syphilis. At present, the serological diagnosis methods of syphilis can be divided into two categories, namely non-treponemal (antigen) serum method...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/531G01N33/571
Inventor 肖洪武
Owner 肖洪武
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