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Method for determining contents of biological marker and DNA through direct-reading portable glucometer

A biomarker and blood glucose meter technology, which is applied in measuring devices, biological testing, instruments, etc., can solve the problem of less blood collection, and achieve the effects of sensitive detection, low analysis cost, and simple operation

Inactive Publication Date: 2013-10-16
HUAZHONG NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As a small tool for measuring blood sugar content, the blood glucose meter has many advantages such as small blood collection volume (only 2 microliters), fast display results, accurate results, easy operation, and low price. It has been widely used by diabetic patients since its inception. application, but there are few similar instruments in the detection of other diseases

Method used

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  • Method for determining contents of biological marker and DNA through direct-reading portable glucometer
  • Method for determining contents of biological marker and DNA through direct-reading portable glucometer
  • Method for determining contents of biological marker and DNA through direct-reading portable glucometer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Determination of phosphorylated p53 protein (p53 15 )Proceed as follows:

[0029] (1) Immunological reaction: Add 100 μL of magnetic nanocomposite Fe to nine 0.5 mL plastic centrifuge tubes 3 o 4 -p53 15 Ab 1 , and then add 100 μL concentration of 0.2ng / mL, 0.5ng / mL, 1ng / mL, 2ng / mL, 5ng / mL, 10ng / mL, 20ng / mL, 50ng / mL, 100ng / mL to the above nine centrifuge tubes respectively mL of p53 15 , after incubation for 50 minutes, magnetic separation; after washing with phosphate buffer solution for more than 2 times, add 100 μL liposome complex marker p53 15 Ab 2 -liposome, incubated for 40 minutes, magnetically separated;

[0030] (2) Determination: Add 10 μL of 10 mg / mL Triton X-100 to the centrifuge tube to release the glucose molecules from the lumen of the captured liposomes; then remove the immune complex by magnetic separation, and take 20 μL of supernatant droplets On the test strip of the blood glucose meter, directly read the reading of the blood glucose meter,...

Embodiment 2

[0032] Fe used in embodiment 1 3 o 4 -p53 15 Ab 1 preparation of

[0033] 1), 200μL 5.0mg / mL carboxylated magnetic nanoparticles Fe 3 o 4 -COOH, dispersed in 1.0 mL of pH 5.2 sulfo fatty acid methylate containing 400 mM 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride and 100 mM N-hydroxysuccinimide In ester sodium salt buffer solution, magnetically separate after activation for 30min, and wash with phosphate buffer solution several times to remove excess 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride and N - hydroxysuccinimide;

[0034] 2), the Fe 3 o 4 -COOH dispersed in 1.0 mL of 20 μg / mL p53 15 Ab 1 Reacted for 12 h in the medium, washed 3 times with phosphate buffer solution after magnetic separation, and the obtained magnetic nanocomposite Fe 3 o 4 -p53 15 Ab 1 Disperse in phosphate buffer solution containing 1% bovine serum albumin and store at 4°C for later use.

Embodiment 3

[0036] p53 used in Example 1 15 Ab 2 - Preparation of liposome composite markers

[0037] The first step is to prepare biotin-labeled liposomes for embedding glucose: Weigh 124 mg of hydrogenated soybean lecithin, 25 mg of cholesterol, and 6 mg of distearylethanolamine-polyethylene glycol-biotin into a 50 mL round-bottomed flask. Stearyl ethanolamine-polyethylene glycol-biotin molar ratio 159:64:2, dissolved in 15mL mixed solvent, mixed solvent is a mixture of chloroform, isopropyl ether and methanol, chloroform:isopropyl ether:methanol volume ratio=6: 6:1, sonicate under nitrogen protection at 45°C until the mixture is evenly dispersed, then add 5mL of 5mg / mL glucose solution at 45°C to the above mixture and sonicate for 5 minutes to form an emulsion with uniformly dispersed particles; then at 45°C Rotary evaporation under reduced pressure to remove the organic solvent to obtain a gel-like dispersion; the resulting liposome solution was incubated in a water bath at 45°C for...

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Abstract

A method for determining the contents of a biological marker and DNA through a direct-reading portable glucometer is characterized in that a glucose molecule embedded carrier liposome, hollow metal ball, porous carbon ball, porous silicon ball or polymer is introduced into the surface of a sensor as a composite marker through a sandwich immunization reaction or a DNA reaction, so the detection of the biological marker or DNA is converted into the detection of a final product glucose, the contents of the biological marker and DNA for detection are in direct proportion to the content of the embedded glucose according to the sandwich immunization analysis principle, and the contents of the biological marker and DNA can be obtained by detecting the concentration of glucose through the glucometer. The glucometer is developed into a portable and universal biological marker and DNA tester, and the glucometer detection technology based biological marker and DNA analysis new method is constructed, and can be used for the real-time, online, simple and sensitive determination of various biological markers and DNA. An apparatus used in the method is simple, so the analysis cost is low.

Description

technical field [0001] The invention relates to a method for measuring biomarker and deoxyribonucleic acid (DNA) content by using a direct-reading portable blood glucose meter. It specifically relates to the preparation method of composite nanocarriers embedding glucose molecules, the conversion of biomarkers or DNA detection into the detection of final product glucose, and the analysis method for measuring various biomarkers and DNA content using blood glucose meter detection technology. Background technique [0002] Biomarkers are signaling indicators of abnormalities at different biological levels due to the influence of environmental pollutants before the organism is seriously damaged. It can provide early warning of severe toxic damage, and can be used for disease diagnosis, judgment of disease stage, or to evaluate the safety and effectiveness of new drugs or new treatments in the target population, and its content is an important indicator for objectively evaluating i...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N33/531
Inventor 杜丹
Owner HUAZHONG NORMAL UNIV
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