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Quantitative fructose assay kit and application thereof as well as quantitative seminal plasma fructose assay method

A quantitative detection and kit technology, which is applied in the preparation of test samples, biochemical equipment and methods, and the determination/inspection of microorganisms, etc., can solve the problems of low sensitivity of resorcinol, operators, environmental hazards, and clinical use. Convenience and other issues, to achieve the effect of high sensitivity, simple operation, convenient clinical routine application and promotion

Inactive Publication Date: 2011-05-18
BRED LIFE SCI TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The indole method is the methodology recommended by the World Health Organization, but it needs to use concentrated hydrochloric acid during the operation, which will cause harm to the operator and the environment; the resorcinol method has low sensitivity and needs to use concentrated hydrochloric acid, which will cause harm to the operator and the environment. Hazardous, and incubating at 90°C for color development, inconvenient for clinical use

Method used

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  • Quantitative fructose assay kit and application thereof as well as quantitative seminal plasma fructose assay method
  • Quantitative fructose assay kit and application thereof as well as quantitative seminal plasma fructose assay method
  • Quantitative fructose assay kit and application thereof as well as quantitative seminal plasma fructose assay method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] One, prepare the fructose quantitative detection kit of the present embodiment 1

[0033] 1. Prepare deproteinized solution A:

[0034] 1) Weigh 6.183 grams of boric acid, put it in a beaker, add an appropriate amount of purified water and stir to dissolve, then dilute to 1L with purified water, mix well and store. (The concentration of boric acid is 0.1mol / L.)

[0035] 2. Preparation of deproteinized solution B:

[0036] Weigh 4 grams of sodium hydroxide, put it in a beaker, add an appropriate amount of purified water and stir to dissolve, then dilute to 1 L with purified water, mix well and store. (Sodium hydroxide concentration is 0.1mol / L.)

[0037] 3. Preparation of reagent 1:

[0038] (1) Weigh 17.728 grams of disodium hydrogen phosphate 12 water, 6.94 grams of potassium dihydrogen phosphate, add an appropriate amount of purified water and stir to dissolve, then adjust the pH value to 6~ with concentrated hydrochloric acid or concentrated sodium hydroxide solu...

Embodiment 2

[0083] One, prepare the fructose quantitative detection kit of the present embodiment 2

[0084] 1. Prepare deproteinized solution A:

[0085] Take an appropriate amount of purified water and add it to the beaker, measure 0.544ml of sulfuric acid, pour it slowly along the wall of the beaker and keep stirring to mix, then dilute to 1L with purified water, mix well and store. (The concentration of sulfuric acid is 0.001mol / L.)

[0086] 2. Preparation of deproteinized solution B:

[0087] Weigh 0.106 g of sodium carbonate, put it in a beaker, add an appropriate amount of purified water and stir to dissolve, then dilute to 1 L with purified water, mix well and store. (Sodium carbonate concentration is 0.001mol / L.)

[0088] 3. Preparation of reagent 1:

[0089] (1) First prepare 0.01mol / L citric acid buffer solution: weigh 2.1014 grams of citric acid in 1 water, add an appropriate amount of purified water and stir to dissolve, then adjust the pH value to 6-7 with concentrated s...

Embodiment 3

[0133] One, prepare the fructose quantitative detection kit of the present embodiment 3

[0134] 1. Prepare deproteinized solution A:

[0135] Measure 82.117ml of hydrochloric acid, put it in a beaker, add an appropriate amount of purified water and stir to dissolve, then dilute to 1L with purified water, mix well and store. (The concentration of hydrochloric acid is 1mol / L.)

[0136] 2. Preparation of deproteinized solution B:

[0137] Weigh 84.01 g of sodium bicarbonate, put it in a beaker, add an appropriate amount of purified water and stir to dissolve, then dilute to 1 L with purified water, mix well and store. (The concentration of sodium bicarbonate is 1mol / L.)

[0138] 3. Preparation of reagent 1:

[0139] (1) Measure 57.234ml of acetic acid, add an appropriate amount of purified water and stir to dissolve, then adjust the pH value to 6-7 with concentrated hydrochloric acid or concentrated sodium hydroxide solution under the monitoring of a pH meter, to obtain the ...

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Abstract

The invention discloses a quantitative fructose assay kit which comprises inorganic acid deproteinized extract A, inorganic base deproteinized extract B, fructose calibration solution, a reagent 1 containing 0.001-0.1mol / L adenosine triphosphate sodium salt, a reagent 2 containing 1-100KU / L hexokinase and 1-100KU / L glucose-6-phosphate dehydrogenase, and a reagent 3 containing 0.001-0.1mol / L nicotinamide adenine dinucleotide. The seminal plasma fructose assay method comprises the following steps: respectively adding the reagent 1 and the reagent 2 to deproteinized seminal plasma and the fructose calibration solution, and mixing uniformly; reacting at the temperature of 10-40 DEG C for 5-120 minutes, then reading the absorbance respectively at the wavelength of 280-400nm; adding the reagent 3 respectively, and mixing uniformly; reacting under the same conditions and reading the absorbance; and calculating the difference between the absorbance read at the first time and the absorbance read at the second time, and comparing or calculating the absorbance of a seminal plasma specimen and the fructose calibration solution to obtain the concentration of the seminal plasma fructose. The kit and the method can be used for quantitative determination of fructose in sera, plasma, body fluid, food and solid extracting solution, the methodology is special, unique, clean and environment-friendly, manual operation and automatic batch assay can be realized, and the kit and the method are easy to popularize and apply clinically.

Description

technical field [0001] The invention relates to a fructose quantitative detection kit and its application, and to a method for using the kit to detect the concentration of seminal plasma fructose in human seminal plasma specimens in vitro. Background technique [0002] Male bilateral ejaculatory duct obstruction, congenital absence of seminal vesicles or dysplasia, fructose was negative. The fructose content decreases in cases of vesiculitis, incomplete ejaculation, or excessive ejaculation. Fructose combined with other detection indicators (such as seminal plasma neutral α-glucosidase) can locate the obstruction site in patients with obstructive azoospermia. Seminal berry fructose is an evaluation function index of seminal vesicle secretion function. The level of testosterone affects the secretion of fructose in the seminal vesicles, and insufficient androgen can cause a decrease in fructose content. Decreased seminal vesicle secretion can lead to decreased semen volume....

Claims

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Application Information

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IPC IPC(8): C12Q1/48C12Q1/32G01N21/31G01N1/34
Inventor 邓少君程锦军王铮铮庄学敏李巍钟彩颜王希上杨红芳
Owner BRED LIFE SCI TECH
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