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Seminal plasma neutral alpha-glucosidase assay kit

A glucosidase and kit technology, applied in the biological field, can solve the problems of complicated operation steps, difficult to popularize and apply, and achieve the effects of simplifying operation steps, reducing manual operations and reducing costs

Inactive Publication Date: 2019-07-05
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

Above-mentioned prior art reagent is of a great variety, and operation step is complicated, is difficult for popularization and application, and needs to be further improved and enhanced

Method used

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  • Seminal plasma neutral alpha-glucosidase assay kit
  • Seminal plasma neutral alpha-glucosidase assay kit
  • Seminal plasma neutral alpha-glucosidase assay kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Composition and preparation of embodiment 1 kit

[0051] Preparation of substrate A: Add 7.5g of dipotassium hydrogen phosphate and 4.5g of potassium dihydrogen phosphate into a 100mL mixing bottle, stir for 20 minutes and mix well, check the pH value of the solution with a pH meter to be 7.2-7.6, add p-nitrophenol- Maltoside 0.07g.

[0052] Preparation of substrate B: Add 7.5g of dipotassium hydrogen phosphate and 4.5g of potassium dihydrogen phosphate into a 100mL mixing bottle, stir for 20 minutes and mix well, check the pH value of the solution with a pH meter to be 7.2-7.6, add β-glucosidase 0.9KU.

[0053] Preparation of stop solution: Accurately weigh 100ml of purified water and 12g of sodium hydroxide, then put them into a mixing bottle, shake or stir to dissolve.

[0054] Preparation of calibration solution: Accurately weigh 1000ml of purified water, 0.022g of p-nitrophenol, and 4g of sodium hydroxide, and then put them into the mixing bottle.

[0055] Prepa...

Embodiment 2

[0057] Embodiment 2 Kit operation and result calculation

[0058] Dilute the calibration solution down to 3 gradients, serial dilution method: take 3 centrifuge tubes, add 300 μL of stop solution to each tube, add 300 μL of calibration solution to the first tube, mix well and take 300 μL from it to the second tube, and so on Dilute to the third tube.

[0059] Another stop solution was taken as the zero calibration solution, and 270 μL of each calibration solution was placed in a microwell plate, and the absorbance value was read colorimetrically at a wavelength of 405 nm, and a quadratic curve was drawn using the absorbance value of the calibration solution and the corresponding concentration of neutral α-glucosidase as a calibration curve.

[0060] Dilute the seminal plasma sample to be tested 1:10: take 1 volume of seminal plasma and 9 volumes of diluent and mix evenly (for example, take 20 μL of seminal plasma and 180 μL of diluent), complete the operation in a microwell p...

Embodiment 3

[0062] Example 3 The kit is integrated into the fully automatic biochemical platform

[0063] With the iMagic-M7 automatic biochemical analyzer (product registration number: Guangdong Food and Drug Administration (Zhun) Zi 2014 No. 2401079) produced by Shenzhen Kuber Biotechnology Co., Ltd. as the experimental platform, this kit can be fully integrated until the device operates autonomously.

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Abstract

The invention provides a seminal plasma neutral alpha-glucosidase assay kit. The kit adopts a double-substrate mode. An enzyme decomposition method is divided into two steps. The seminal plasma blankmode is changed. A glycosidase inhibitor is removed. Sodium hydroxide is used to stop reaction. The reaction mode is adjusted. The reaction cost is reduced. The reaction time is shortened. The resultis accurate and stable. The seminal plasma neutral alpha-glucosidase assay kit has a broad application prospect and a great market value.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a test kit for seminal plasma neutral α-glucosidase. Background technique [0002] Neutral alpha-glucosidase (neutral alpha-glucosidase) is an indicator of secretory function of the epididymis. It is more specific and sensitive to the epididymis than L-carnitine and glycerophosphocholine. Combined with hormones and other testicular indicators, it has a good diagnostic value for distal vas deferens obstruction. The detection of fructose combined with neutral α-glucosidase can greatly improve its value in the diagnosis of obstructive azoospermia. [0003] The principle of the seminal plasma neutral α-glucosidase assay recommended on pages 113-115 of the "Human Semen Examination and Processing Laboratory Manual" (5th Edition) compiled by the World Health Organization (WHO) is: glucosidase will The synthesized glucopyranoside substrate was converted to p-nitrophenol, which turned yellow...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31
CPCG01N21/31
Inventor 李明磊傅剑华蔡华曾周祥蔡栋庄兴华邹玮
Owner 深圳华康生物医学工程有限公司