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Glucose detection method, kit and application thereof

A detection method, glucose technology, applied in the field of glucose detection, can solve the problems of low accuracy, poor specificity, influence, etc., and achieve the effect of high sensitivity and strong specificity

Pending Publication Date: 2019-03-05
KANGMA SHANGHAI BIOTECH LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the aforementioned method has disadvantages such as poor specificity, low accuracy, and easy influence of other small molecules in the body fluid.

Method used

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  • Glucose detection method, kit and application thereof
  • Glucose detection method, kit and application thereof
  • Glucose detection method, kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0110] In the present invention, the preparation method of the cell extract is not limited, and a preferred preparation method includes the following steps:

[0111] (i) providing cells;

[0112] (ii) washing the cells to obtain washed cells;

[0113] (iii) performing cell disruption treatment on the washed cells to obtain a crude cell extract;

[0114] (iv) performing solid-liquid separation on the crude cell extract to obtain the liquid part, which is the cell extract.

[0115] In the present invention, the solid-liquid separation method is not particularly limited, and is preferably centrifugation.

[0116] In the present invention, the centrifugation conditions are not particularly limited, and the centrifugation conditions are 5000-100000g, preferably 8000-30000g.

[0117] In the present invention, the centrifugation time is not particularly limited, and the centrifugation time is 0.5min-2h, preferably 20min-50min.

[0118] In the present invention, the temperature of...

Embodiment approach

[0192] As a specific embodiment, the present invention provides a test kit for glucose concentration detection, comprising:

[0193] (k1) a first container, and a cell extract located in the first container;

[0194] (k2) a second container, and an RNA polymerase located in the second container;

[0195] (k3) a third container, and a DNA template located in the third container;

[0196] (k4) a fourth container, and a substrate for synthesizing RNA located in the fourth container;

[0197] (k5) the 5th container, and the substrate that is positioned at the 5th container and is used for synthesizing protein;

[0198] (k6) a sixth container, and the phosphate located in the sixth container;

[0199] (kt) label or instructions.

[0200] (k7) a seventh container, and magnesium ions located in the seventh container;

[0201] (k8) an eighth container, and potassium ions located in the eighth container;

[0202] (k9) a ninth container, and a buffer located in the ninth container...

Embodiment 1

[0221] Embodiment 1: Determination of Luminous Intensity-Glucose Concentration Standard Curve of the present invention

[0222] 1.1 Provide or prepare an in vitro protein synthesis system;

[0223] 1.2 Add different concentrations of glucose (1.1-12.8mM) into the in vitro protein synthesis system as the energy source for the system to synthesize the reporter protein;

[0224] 1.3 At room temperature (20-25°C), incubate the above in vitro protein synthesis system for about 30 minutes, and the glucose in the system has been completely reacted after detection;

[0225] 1.4 Add an equal volume of substrate luciferine and the above-mentioned reaction samples to a 96-well white plate or a 384-well white plate, and immediately place it on an Envision 2120 multifunctional microplate reader (Perkin Elmer) for reading to detect firefly luciferase activity , the relative light unit value (RLU) is used as the activity unit;

[0226] 1.5 Take the glucose concentration as the abscissa and...

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Abstract

The invention provides a glucose detection method. The glucose detection method comprises the following steps: (1) preparing: preparing an in-vitro biological synthetic system; (2) adding: adding a sample into the synthetic system in step (1); (3) synthesizing: starting the in-vitro biological synthetic reaction of a target object; (4) detecting: detecting a signal intensity of the synthesized target object in the synthetic system; and (5) calculating: calculating a glucose concentration or content of a to-be-detected sample according to a signal intensity-glucose concentration relation curve.The invention also provides a glucose detection kit adopting the detection method stated in a first aspect and an application of the glucose detection kit in detecting blood glucose and urine glucose. Compared with a glucose oxidase electrode method in the prior art, the glucose detection method has the advantages of high specificity, high sensitivity, less external interference, ease in high-flux and big data measurement and the like.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a glucose detection method, a kit and an application thereof. Background technique [0002] Glucose is an important source of energy in the human body, and its concentration in the blood is tightly regulated. Changes in blood glucose concentration can reflect the health status of the human body, especially the physical condition of diabetic patients. Prolonged high blood sugar or frequent fluctuations in blood sugar concentration can easily cause various complications. Generally speaking, the normal blood sugar value is as follows: the normal value of fasting blood sugar is 4.0-6.1mmol / L, and the blood sugar range of 1 hour after a normal meal is 6.7-9.4 mmol / L, and the blood sugar range of two hours is 3.9-7.8mmol / L. Fasting whole blood glucose ≥ 6.7mmol / L, 2 repeated measurements can be diagnosed as diabetes. Only when the blood sugar exceeds a certain concentration range, more s...

Claims

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Application Information

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IPC IPC(8): C12Q1/54C12Q1/66G01N21/64
CPCG01N21/6428C12Q1/54C12Q1/66
Inventor 郭敏徐开雷先德于雪
Owner KANGMA SHANGHAI BIOTECH LTD
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