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Detection method of HPV-E6 proteins

An HPV-E6 and detection method technology, applied in the field of biochemistry, can solve the problems of cumbersome operation, inability to perform batch sample analysis, etc., and achieve the effects of good repeatability, simple operation and high accuracy

Inactive Publication Date: 2014-10-01
沈萍萍 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods have the disadvantages of cumbersome operation, special equipment, sample pretreatment, batch sample analysis and automatic biochemical analyzer detection.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Prepare HPV-E6 protein reaction reagent: take 50ml of 5mmol / L phosphate buffer saline as the base, add 0.1% sodium azide, 0.1% magnesium chloride, 0.1% electrolyte sodium chloride and 20% polyethylene Diol 2000 mixed;

[0024] Preparation of anti-HPV-E6 protein antibody reagent: take 20ml goat anti-human HPV-E6 protein antiserum, use it as the base, add 0.001% of its weight butylated hydroxyanisole and 20% of magnesium chloride and mix;

[0025] Prepared by mixing HPV-E6 protein reagent and anti-HPV-E6 protein antibody reagent at a volume ratio of 3:1;

[0026] Prepare a constant value calibration solution for serotypes: take 50ml of diglycine with a concentration of 20mmol / L, add 1% of its volume sodium azide at a concentration of 0.01 to 10mmol / L, 5% disodium edetate, The concentration is 0.1~40mmol / L;

[0027] Measure the absorbance value of the HPV-E6 protein reactant and anti-HPV-E6 protein antibody reagent mixture at a wavelength of 340nm, compare it with a sero...

Embodiment 2

[0029] Preparation of HPV-E6 protein reaction agent: take 60ml of tris buffer solution with a concentration of 40mmol / L, use it as the base, add 0.5% p-hydroxybenzoic acid, 0.3% ethylene glycol, and 5% electrolyte chloride by weight Sodium and 25% polyethylene glycol 4000 are mixed, add buffer solution weight base 0.1% nonionic surfactant and 2% concentration in addition and be 0.01mmol / L hexamethylene dimethylamine bromide;

[0030] Preparation of anti-HPV-E6 protein antibody reagent: take 30ml of horse anti-human HPV-E6 protein antiserum, use it as the base, add 1% dilauryl thiodipropionate by weight and 25% ethylene glycol to mix;

[0031] Prepared by mixing HPV-E6 protein reagent and anti-HPV-E6 protein antibody reagent at a volume ratio of 10:1;

[0032] Prepare a serotype constant value calibration solution: take 60ml of bisglycine with a concentration of 30mmol / L, add its volume of 2% p-hydroxybenzoic acid at a concentration of 1mmol / L, and 8% magnesium chloride at a co...

Embodiment 3

[0035] Preparation of HPV-E6 protein reaction reagent: take 70ml of buffer solution with a concentration of 80mmol / LPIPES, use it as the base, add 1% phenol, 0.5% trehalose, 10% electrolyte sodium chloride and 30% polyethylene glycol 6000 to mix ;

[0036] Preparation of anti-HPV-E6 protein antibody reagent: take 40ml of mouse anti-human HPV-E6 protein antiserum, use it as the base, add 2% of its weight of BHA and 30% of trehalose to mix;

[0037] Prepared by mixing HPV-E6 protein reagent and anti-HPV-E6 protein antibody reagent at a volume ratio of 30:1;

[0038] Prepare a serotype constant value calibration solution: take 70ml of glycine with a concentration of 40mmol / L, add 3% phenol by volume, the concentration is 3mmol / L, and 15% glycerol, the concentration is 10mmol / L;

[0039] Measure the absorbance value of the HPV-E6 protein reactant and anti-HPV-E6 protein antibody reagent mixture at a wavelength of 340nm, compare it with a serotype constant value calibration soluti...

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PUM

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Abstract

The invention discloses a detection method of HPV-E6 proteins. The method comprises the steps that an HPV-E6 protein reactant is prepared, an HPV-E6 protein antibody reagent is prepared, the reactant and the antibody reagent are mixed, the light absorption value of the reactant and the antibody reagent is compared with that of constant-value standard liquid, and the HPV-E6 protein content in a sample is calculated out. The method has the advantages that the sample is free of dilution, operation is easy, the accuracy is high, repeatability is good, the capacity of resisting disturbance is high, and the method is applicable to fully automatic biochemical analyzers of various types.

Description

technical field [0001] The invention belongs to the field of biochemistry, and in particular relates to a method for detecting HPV-E6 protein. Background technique [0002] HPV virus (abbreviation for human papillomavirus) is a kind of papillomavacuolating virus A belonging to the Papovaviridae family. It is a spherical DNA virus that can cause squamous epithelial proliferation of human skin and mucous membranes. At present, more than 130 species have been isolated. This virus only invades humans and is not pathogenic to other animals. Once infected, the human body will carry it for life. Different types cause different clinical manifestations, about 35 types are related to reproductive tract infection, about 20 types are related to cancer, and there are 13 types of high-risk HPV related to cervical cancer: 16, 18, 31, 33, 35 , 39, 45, 51, 52, 56, 58, 59, 68; 16 and 18 are more common. HPV-E6 protein is a protein specifically expressed by types 16 and 18, and is used as an...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N21/33G01N1/38
Inventor 沈萍萍张帅陈霆
Owner 沈萍萍
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