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Detection reagent and detection method of alpha-amylase

A technology for detection reagents and detection methods, applied in biochemical equipment and methods, microbiological determination/inspection, etc., can solve the problems of unconditional purchase and use, high requirements for supporting conditions, complicated operation, etc., and achieve simple operation and low cost , the effect of simple operation

Active Publication Date: 2014-07-02
HUNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, these traditional methods for detecting α-amylase require large and expensive instruments, such as large-scale automatic biochemical analyzers, to calculate the concentration of α-amylase by measuring the absorbance value of the hydrolysis end product
Due to the high price and complicated operation of large-scale automatic biochemical instruments, operators need relevant professional knowledge and training, and the equipment has high requirements for supporting conditions and requires regular maintenance by professionals. Therefore, primary medical institutions or family individuals have no conditions to purchase and use.

Method used

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  • Detection reagent and detection method of alpha-amylase
  • Detection reagent and detection method of alpha-amylase
  • Detection reagent and detection method of alpha-amylase

Examples

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Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Detection of different concentrations of α-amylase

[0035] A detection reagent for α-amylase of the present invention is composed of malt pentose, α-glucosidase, calcium chloride and disodium hydrogen phosphate-potassium dihydrogen phosphate buffer solution. Among them, the concentration of maltopentose in the buffer is 30mmol / L, the concentration of α-glucosidase in the buffer is 2000U / L, the concentration of calcium chloride in the buffer is 30mmol / L, disodium hydrogen phosphate - The potassium dihydrogen phosphate buffer solution has a pH value of 7.3 and a concentration of 10 mmol / L, that is, includes 0.0408 g of disodium hydrogen phosphate and 0.2507 g of potassium dihydrogen phosphate.

[0036] The above-mentioned α-amylase detection reagent is obtained by the following preparation method:

[0037] (1) Prepare maltpentose solution, α-glucosidase solution and calcium chloride solution respectively, and the solvents are all disodium hydrogen phosphate...

Embodiment 2

[0044] Embodiment 2: specificity investigation

[0045] The detection reagent of α-amylase used in this example is the same as that in Example 1.

[0046] Prepare 50nmol / L standard solutions of different substances (used to replace the α-amylase solution), and the different substances specifically investigated in this example include thrombin, hemoglobin, human serum albumin (HSA) and C-reactive protein (CRP) .

[0047] Mix 50nmol / L thrombin solution, hemoglobin solution, HSA solution, CRP solution and α-amylase solution (AMS, that is, 540U·L -1 ) were added to the reaction system under the same conditions as in Example 1 (i.e., the operating temperature was 37°C, and the catalytic reaction time was 15 minutes). After the reaction was complete, 4 μL of the reaction product was taken for testing with a blood glucose meter, and the reading of the blood glucose meter was recorded after 25 seconds. , the result is as image 3 shown. From image 3 It can be seen from the det...

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Abstract

The invention discloses a detection reagent and a detection method of alpha-amylase. The detection reagent comprises 10mmol / L-40mmol / L of malt pentose, 1000U / L-5000U / L of alpha-glucosidase, 10mmol / L-30mmol / L of calcium chloride and 1mmol / L-50mmol / L of a buffer solution. The detection method comprises the following steps: adding an alpha-amylase solution into the detection reagent and performing catalytic reaction to obtain a reaction product; detecting the reaction product by virtue of a portable glucose detection instrument; recording the readings of the detection instrument; and calculating the concentration of the alpha-amylase solution. The detection reagent can quickly and quantitatively detect the alpha-amylase; the detection method is easy and convenient to operate, quick, high in sensitivity and good in selectivity, the used instrument is portable, and the alpha-amylase can be quickly detected by people who do not accept professional training.

Description

technical field [0001] The invention belongs to the field of biotechnology and relates to a detection reagent and a detection method, in particular to a detection reagent for α-amylase and a method for rapidly and quantitatively detecting α-amylase by using a portable blood glucose meter or other glucose detectors. Background technique [0002] α-amylase (α-1,4-D-glucose-glucoside hydrolase) can cut α-1,4 glucosidic bonds in starch, glycogen, oligomeric or polysaccharide molecules to produce maltose, oligomeric sugar and glucose etc. Alpha-amylase is synthesized by secretory epithelial cells in the salivary glands and pancreas. In cases of mumps, especially acute pancreatitis, the concentration of α-amylase in blood and urine increases significantly. Some acute abdomen, such as acute appendicitis, intestinal obstruction, pancreatic cancer, perforated ulcer disease, etc. can cause serum α-amylase to increase, so the detection of α-amylase has important clinical significance...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/40C12Q1/34
Inventor 王青王柯敏王辉羊小海翦立新
Owner HUNAN UNIV
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