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INS5 insulin kit based on aptamer fluorescent probe and detection method of INS5 insulin

A nucleic acid aptamer and fluorescent probe technology, applied in the field of medical testing and determination, can solve the problems of operation technical errors, many interference factors, poor repeatability, etc., and achieve the effects of high repeatability, strong specificity and convenient transportation.

Inactive Publication Date: 2017-03-22
LIUZHOU LIJIE TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention is to provide a nucleic acid aptamer fluorescent probe-based insulin INS kit and its detection for the existing deficiencies such as easy to produce operational technical errors, poor repeatability, many interference factors, cumbersome operations, and high testing costs. method

Method used

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  • INS5 insulin kit based on aptamer fluorescent probe and detection method of INS5 insulin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] After dissolving and preparing the required reagent components according to Example 1 in Table 1, divide into bottles and freeze-dry to make a dry powder reagent; before use, add ultrapure water and use after reconstitution. Three parallels were set for each sample, and the detection steps were as follows:

[0033] (1) Blood sample lysing: Mix heparin anticoagulated whole blood and erythrocyte lysate at a volume ratio of 1:0.5, let stand for 30 minutes, then centrifuge at a medium speed for 7 minutes, and collect the supernatant;

[0034] (2) Mixed egg incubation: take 20 μl of the supernatant obtained in step 1) and 45ul of the insulin aptamer fluorescent probe reagent obtained by dissolving the insulin aptamer fluorescent probe in 0.2M phosphate buffer, mix, and incubate eggs at room temperature for 5 minutes. get the test solution;

[0035] (3) Fluorescence detection: 50ul of the test solution obtained in the detection step 2) of the fluorescence detector, after the...

Embodiment 2

[0039] After dissolving and preparing the required reagent components according to Example 2 in Table 1, divide into bottles and freeze-dry to make a dry powder reagent; before use, add ultrapure water and use after reconstitution. Three parallels were set for each sample, and the detection steps were as follows:

[0040] (1) Blood sample lysis: Mix peripheral blood and red blood cell lysate at a volume ratio of 1:2.5, let stand for 5 minutes, then centrifuge at a medium speed for 6 minutes, and collect the supernatant;

[0041] (2) Mixed egg incubation: Take 50μl of the supernatant obtained in step 1) and 30ul of

[0042] The insulin aptamer fluorescent probe reagent obtained by dissolving the insulin aptamer fluorescent probe in 0.2M phosphate buffer solution was mixed, and the eggs were incubated at room temperature for 6 minutes to obtain the test solution;

[0043] (3) Fluorescence detection: 50ul of the test solution obtained in the detection step 2) of the fluorescence...

Embodiment 3

[0046] After dissolving and preparing the required reagent components according to Example 3 in Table 1, divide into bottles and freeze-dry to make a dry powder reagent; before use, add ultrapure water and use after reconstitution. Three parallels were set for each sample, and the detection steps were as follows:

[0047] (1) Blood sample lysis: Mix peripheral blood and red blood cell lysate at a volume ratio of 1:3.5, let stand for 10 minutes, then centrifuge at a medium speed for 5 minutes, and collect the supernatant;

[0048] (2) Mixed egg incubation: Take 75μl of the supernatant obtained in step 1) and 45ul of

[0049] The insulin aptamer fluorescent probe reagent obtained by dissolving the insulin aptamer fluorescent probe in 0.2M phosphate buffer solution was mixed, and the eggs were incubated at room temperature for 7 minutes to obtain the test solution;

[0050] (3) Fluorescence detection: 50ul of the test solution obtained in the detection step 2) of the fluorescenc...

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Abstract

The invention relates to an insulin INS kit based on an aptamer fluorescent probe, further relates to a method for measuring the insulin INS concentration, measuring reagent composition and components, and belongs to the technical field of medical examination and measurement. The kit mainly comprises red cell lysate, a phosphate buffer solution (PBS), an insulin (INS) standard substance and the insulin (INS) aptamer fluorescent probe. The concentration of insulin (INS) is measured through blood sample pyrolysis, ovum cultivation processing and fluorospectro photometer detection. The kit and the method have the advantages that sample treatment is easy, operation is easy, detection time is short, detection specificity is high, sensitivity is high, and detection results are high in repeatability.

Description

technical field [0001] The invention belongs to the technical field of medical testing and determination, in particular to an insulin kit based on a nucleic acid aptamer fluorescent probe INS5 and a detection method thereof. Background technique [0002] Insulin is a protein hormone secreted by the islet β cells in the pancreas when stimulated by endogenous or exogenous substances such as glucose, lactose, ribose, arginine, and glucagon. Insulin is the only hormone in the body that lowers blood sugar, and at the same time promotes the synthesis of glycogen, fat, and protein. Exogenous insulin is mainly used in the treatment of diabetes. [0003] The biosynthesis rate of insulin is affected by the concentration of plasma glucose. When the concentration of blood glucose rises, the content of proinsulin in β cells increases and the synthesis of insulin accelerates. Insulin is synthesized in the beta cells of the pancreas. Insulin has a molecular weight of 5700 and consists o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64G01N33/66G01N33/68G01N33/74
CPCG01N21/6486G01N21/6408G01N33/66G01N33/68G01N33/6893G01N33/74G01N2333/62G01N2800/042
Inventor 陈东
Owner LIUZHOU LIJIE TECH
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