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Homocysteine kit based on aptamer fluorescence probe HCy2 and detection method thereof

A homocysteine ​​and fluorescent probe technology, applied in the field of homocysteine ​​HCy kits, can solve the problems of operation technical error, poor repeatability, and high test cost, and achieve high repeatability, convenient transportation, Sensitive effect

Inactive Publication Date: 2015-05-06
深圳市一道生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The purpose of the present invention is to provide a homocysteine ​​HCy reagent based on a nucleic acid aptamer fluorescent probe for the existing deficiencies such as easy to produce operational technical errors, poor repeatability, many interference factors, cumbersome operations, and high testing costs. Box and detection method thereof

Method used

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  • Homocysteine kit based on aptamer fluorescence probe HCy2 and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] 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:

[0035] (1) Lysis of blood samples: 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;

[0036] (2) Mixed egg incubation: take 20 μl of the supernatant obtained in step 1) and 45ul of the homocysteine ​​nucleic acid aptamer fluorescent probe reagent obtained by dissolving the homocysteine ​​nucleic acid aptamer fluorescent probe in 0.2M phosphate buffer Mix and incubate the eggs at room temperature for 5 minutes to obtain the test solution;

[0037] (3) Fluorescence detection: 50ul of the test solution obtained in the detecti...

Embodiment 2

[0041] 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:

[0042] (1) Lysis of blood samples: Mix peripheral blood and erythrocyte 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;

[0043] (2) Mixed egg incubation: take 50 μl of the supernatant obtained in step 1) and 30ul of the homocysteine ​​nucleic acid aptamer fluorescent probe reagent obtained by dissolving the homocysteine ​​nucleic acid aptamer fluorescent probe in 0.2M phosphate buffer Mix and incubate the eggs at room temperature for 6 minutes to obtain the test solution;

[0044] (3) Fluorescence detection: 50ul of the test solution obtained in the detection step 2) of the f...

Embodiment 3

[0047] 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:

[0048] (1) Lysis of blood samples: Mix peripheral blood and erythrocyte 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;

[0049] (2) Mixed egg incubation: take 75 μl of the supernatant obtained in step 1) and 45ul of the homocysteine ​​nucleic acid aptamer fluorescent probe reagent obtained by dissolving the homocysteine ​​nucleic acid aptamer fluorescent probe in 0.2M phosphate buffer Mix and incubate the eggs 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 ...

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Abstract

The invention relates to a homocysteine Hcy kit based on an aptamer fluorescence probe; at the same time, the invention also relates to a method for determination of the concentration of homocysteine Hcy, and a determination reagent composition and components, and belongs to the technical field of medical examination and determination. The kit comprises the main components: an erythrocyte lysate, a phosphate buffer solution PBS, a homocysteine Hcy standard substance, and the homocysteine Hcy aptamer fluorescent probe; through blood sample cracking and mixed egg fertility treatment, and with combination of a fluorescence spectrophotometer for detection, the value of the concentration of the homocysteine Hcy is measured and calculated. The kit has the advantages of simple sample treatment, simple and convenient operation, short detection time, strong detection specificity, high sensitivity, high repeatability of detection results and the like.

Description

technical field [0001] The invention belongs to the technical field of medical examination and measurement, in particular to a homocysteine ​​HCy kit based on a nucleic acid aptamer fluorescent probe and a detection method thereof. Background technique [0002] Homocysteine ​​is a sulfur-containing amino acid and an intermediate product of methionine metabolism in organisms. Compared with cysteine, there is one more methylene group in the side chain. Homocysteine ​​is a risk factor for many diseases. At present, the increase of homocysteine ​​is considered to be the most extensive and independent pathogenic factor of cardiovascular and cerebrovascular diseases caused by atherosclerosis. Homocysteine ​​is also related to various diseases such as neural tube defects, Parkinson's disease, senile dementia, and chronic renal failure, as well as the effects of certain drugs and tumors. The main methods for detecting homocysteine ​​are chromatographic methods, immunological method...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N21/64
CPCG01N21/6402G01N33/6812G01N2800/2871
Inventor 陈燕婷张莘蔓李红梅
Owner 深圳市一道生物科技有限公司
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