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Seminal plasma neutral alpha-glucosidase quantitative detecting reagent, kit and preparing method thereof

A technology for glucosidase and quantitative detection, which is used in biochemical equipment and methods, and the determination/inspection of microorganisms. Effect

Inactive Publication Date: 2005-07-20
深圳华康生物医学工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In addition, since the detection operation process itself is affected by human factors and instruments and equipment, the accuracy of the operation process cannot be monitored.

Method used

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  • Seminal plasma neutral alpha-glucosidase quantitative detecting reagent, kit and preparing method thereof

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Experimental program
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Effect test

Embodiment 1

[0064] Embodiment 1. A seminal plasma neutral α-glucosidase quantitative detection reagent is composed of the following components: (1) acid inhibitor: 100 milliliters of deionized aqueous solution with a sodium lauryl sulfate content of 5 to 15 g; 2) Glycosidase inhibitors: 100 ml of potassium phosphate buffer solution with papaverin content of 5-50 mg; (3) Preferred glycosidase quality control solution: containing 20-500 mg of α-glucosidase and 0.05-0.5 moles of ammonium sulfate / L, 1000 ml of acetate buffer solution with a glycine content of 0.1-1.0 moles; (4) preferred glycoside substrates: 0.5-5.0 millimoles of 4-methylumbelliferone-a-D glucoside, 100 millimoles of D-fructosidase ~500U, 0.5~5g sodium azide dissolved in 1000ml potassium phosphate buffer; (5) Substrate solution: 5~50g sucrose, 10~100mmol sodium chloride, 0.5~5g casein, dissolved Distilled aqueous solution in 1000 ml; (6) stop solution: sodium carbonate 0.05 ~ 0.5 mol / liter deionized aqueous solution; (7) st...

Embodiment 2

[0066] Embodiment 2. A kit made according to Embodiment 1 seminal plasma neutral α-glucosidase quantitative detection reagent, wherein: acid inhibitor 5-15ul, glucosidase inhibitor 2.5×10 -4 ~2.5×10 -3 mg, quality control solution 2~8ul, substrate: 4-methylumbelliferone-a-D glucoside 0.5×10 -4 ~5.0×10 -4 Millimoles, D-fructosidase 0.01~0.05U, substrate solution 50~150ul, stop solution 500~1500ul, one or more deionized aqueous solutions of different concentrations, the amount of each standard is in the microplate reader The upper absorbance does not exceed 3.0.

[0067] The preferred solution of the above kit is: acid inhibitor 10ul, glycosidase inhibitor 9×10 -4 mg, 4-methylumbelliferone-a-D-glucoside 1.85×10 -4 Millimoles, D-fructosidase 0.03U, substrate solution 100ul, stop solution 1000ul, the standard product is standard product 5 and more than one other standard product prepared according to the following method, each standard product is 250ul:

[0068] 1) Preparatio...

Embodiment 3

[0074] Embodiment 3, a preparation method of seminal plasma neutral α-glucosidase quantitative detection reagent, comprising the following steps:

[0075] A, the preparation of acid inhibitor

[0076] 1) Weigh 5 to 15 grams of sodium lauryl sulfate and place it in a beaker, add 80 ml of deionized water, shake and dissolve in a warm bath at 37°C,

[0077] 2) Transfer to a 100ml volumetric flask, dilute to 100ml with deionized water, and store at room temperature;

[0078] B, the preparation of glycosidase inhibitor

[0079] 3), dissolving 5-50mg papaverine in 100ml 0.2mmol / L potassium phosphate buffer solution,

[0080] 4) Aliquot and freeze-dry, store at 4-8°C;

[0081] C. Preparation of substrate

[0082] 5), weigh 0.5~5.0mmol 4-methylumbelliferone-a-D glucoside, 100~500U D-fructosidase, 0.5~5g sodium azide, dissolve in 1000ml of pH6.5, 0.2mol / L potassium phosphate buffer,

[0083] 6) Aliquot and freeze-dry, store at 4-8°C;

[0084] D. Preparation of stop solution

[...

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Abstract

The present invention discloses one kind of quantitative detecting reagent for neutral alpha-glucosidase in seminal plasma. The detecting reagent includes acid inhibitor, glycosidase inhibitor, quality controlling liquid, substrate, substrate dissolving liquid, terminating liquid, and standard sample. Compounding the substrate can raise the reaction speed greatly, and compounding and detecting the quality controlling liquid can monitor the operation process accurately. The present invention also discloses the reagent kit with the quantitative detecting reagent for neutral alpha-glucosidase in seminal plasma and the preparation process of the detecting reagent.

Description

technical field [0001] The invention belongs to the detection field of biochemical enzyme kits, and is particularly suitable for the quantitative detection of seminal plasma neutral alpha-glucosidase. Background technique [0002] In the clinical and theoretical research of male infertility, how to evaluate the secretory function of the epididymis is a very important and arduous task. The early research indexes of epididymis secretory function included L-carnitine and glycerophosphocholine, and then α-glucosidase was included in the evaluation indexes. The latest research results show that seminal plasma neutral α-glucosidase is the best known index for evaluating epididymis secretory function, and its specificity is better than that of L-carnitine, glycerophosphocholine and α-glucosidase. It can be used for clinical diagnosis and prognosis monitoring of azoospermia, vas deferens obstruction, impaired epididymis function, etc. [0003] The quantitative detection methodolog...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/25
Inventor 傅剑华刘瑜胡家纯何林何小红
Owner 深圳华康生物医学工程有限公司
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