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Kit for separating Escherichia coli in urine and preparation method thereof

A technology of Escherichia coli and a kit, which is applied in the biological field, can solve the problems of high cost of antibodies, and achieve the effects of low cost, improved sensitivity, and controllable quality

Pending Publication Date: 2022-07-29
南昌大学第一附属医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In recent years, pathogenic microorganism detection methods based on immunology and molecular biology have developed rapidly, but each has certain defects, such as expensive antibodies, false positive amplification, etc.

Method used

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  • Kit for separating Escherichia coli in urine and preparation method thereof
  • Kit for separating Escherichia coli in urine and preparation method thereof
  • Kit for separating Escherichia coli in urine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] A test kit and preparation method for separating Escherichia coli in urine are carried out according to the following steps:

[0041] (1) Aspirate 1 mL of magnetic nano-beads (10 mg / mL), add it to 9 mL of sterile PBS solution of pH 7.4, wash the magnetic nano-beads under the action of an external magnetic field, discard the washing solution, and repeat the washing steps in total. 3 times, resuspend the washed magnetic nanobeads in 9 mL of sterile PBS solution;

[0042] (2) Weigh 2.9 mg of EDC and dissolve in 290 μL of sterile PBS, weigh 3.26 mg of NHSS and dissolve in 326 μL of sterile PBS, add the dissolved EDC and NHSS to the washed nano-magnetic bead solution, and then add Add 9mL sterile PBS, activate at 20°C for 1h;

[0043] The activated magnetic nanobeads were washed three times with sterile PBS and resuspended in 9.8 mL of sterile PBS solution;

[0044] (3) Weigh 2 mg of antimicrobial peptide (polymyxin B sulfate), dissolve it in 200 μL of sterile ultrapure wa...

Embodiment 2

[0047] A test kit and preparation method for separating Escherichia coli in urine are carried out according to the following steps:

[0048] (1) Aspirate 1 mL of magnetic nano-beads (10 mg / mL), add it to 9 mL of sterile PBS solution of pH 7.4, wash the magnetic nano-beads under the action of an external magnetic field, discard the washing solution, and repeat the washing steps in total. 3 times, resuspend the washed magnetic nanobeads in 9 mL of sterile PBS solution;

[0049] (2) Weigh 2.9 mg of EDC and dissolve in 290 μL of sterile PBS, weigh 3.26 mg of NHSS and dissolve in 326 μL of sterile PBS, add the dissolved EDC and NHSS to the washed nano-magnetic bead solution, and then add Add 9mL sterile PBS, activate at 25°C for 2h;

[0050] The activated magnetic nanobeads were washed three times with sterile PBS and resuspended in 9.8 mL of sterile PBS solution;

[0051] (3) Weigh 2 mg of antimicrobial peptide (polymyxin B sulfate), dissolve it in 200 μL sterile ultrapure water...

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Abstract

The invention discloses a kit for separating Escherichia coli in urine and a preparation method, and belongs to the technical field of biology. The method comprises the following steps: coupling nano magnetic beads with antibacterial peptide (polymyxin B sulfate), capturing Escherichia coli in a sample solution by using a nano magnetic bead compound solution modified by the antibacterial peptide, separating and resuspending the captured Escherichia coli from the sample solution under the action of an external magnetic field, and the like. The Escherichia coli captured through magnetic separation can be directly subjected to subsequent analysis; the method has the advantages of high enrichment efficiency, good specificity and low cost, does not need expensive instruments and equipment, and can be popularized and used in primary hospitals.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a kit for separating Escherichia coli in urine and a preparation method. Background technique [0002] Diseases caused by pathogenic microorganisms are diseases with high morbidity and mortality worldwide, causing millions of infections every year and causing major obstacles to socio-economic development. Human infection with bacteria can cause bacteremia, meningitis and other related diseases. Therefore, it is necessary to develop rapid screening methods for pathogenic bacteria. In recent years, the detection methods of pathogenic microorganisms based on immunology and molecular biology have developed rapidly, but they have certain defects, such as high cost of antibodies and false positive amplification. In view of the need to establish an efficient, low-cost, and matrix-resistant detection method, sample pretreatment techniques based on functionalized nanomagnetic be...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/543
CPCG01N33/56916G01N33/54326G01N2333/245
Inventor 许恒毅刘洋肖芳斌魏丹丹徐倩牟丹程南燕张伟
Owner 南昌大学第一附属医院
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