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A kind of polymyxin b magnetic bead, preparation method and application thereof

A polymyxin and magnetic bead technology, applied in the field of nanomaterials and bacteria testing, can solve problems such as low efficiency and false positives, and achieve the effects of strong specificity, high accuracy and good adsorption effect.

Active Publication Date: 2021-11-02
SHAANXI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] In view of the above-mentioned limitations existing in the prior art, the purpose of the present invention is to provide a polymyxin B magnetic bead, preparation method and application thereof, to solve the problem of enrichment of Haemophilus influenzae type b by traditional culture methods, especially trace amounts The problem of low efficiency and false positives during bacterial infection

Method used

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  • A kind of polymyxin b magnetic bead, preparation method and application thereof
  • A kind of polymyxin b magnetic bead, preparation method and application thereof
  • A kind of polymyxin b magnetic bead, preparation method and application thereof

Examples

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preparation example Construction

[0039] The preparation method of polymyxin B magnetic beads of the present invention mainly comprises the following steps:

[0040] Step 1: Polymyxin B pretreatment

[0041] Step 1.1: Benzyl chloroformate, diethyl ether, NaHCO 3The solutions are mixed together in dichloromethane solution to prepare the reaction solution, wherein the mass of benzyl chloroformate: NaHCO 3 The quality of ether: the volume of ether=1:0.5~1:0.5~2.5, preferably 1:0.75:2.5, wherein, the unit of mass is g, and the unit of volume is ml; the reaction solution is added dropwise to polymyxin B , stirred at 25-40°C and 300-500r / min for 2-4h, separated to obtain pre-treated polymyxin B, and protected the amino group of polymyxin B; preferably, polymyxin B The mass ratio of mycocin B to benzyl chloroformate is 3:2;

[0042] Wherein, the separation process of polymyxin B is as follows: adding citric acid and chloroform to the mixed solution, and centrifuging to collect the water phase, which is the primary...

Embodiment 1

[0057] Step 1: Polymyxin B pretreatment

[0058] Step 1.1: Weigh 0.3g of polymyxin B and place it in a 250mL flask, accurately absorb 0.14mL benzyl chloroformate (Cbz-cl) with a concentration of 1mol, 0.5mL ether, and 10% NaHCO3 Dissolve 1.5mL of the aqueous solution in 50mL of dichloromethane to prepare the reaction solution;

[0059] Add the reaction solution dropwise to the flask containing polymyxin B, stir for 2.5 hours at a temperature of 30°C, a rotation speed of 400r / min, and a stirring paddle diameter of 100mm; then add 5mL of 10% lemon to the mixture dropwise. Acid, 20mL chloroform, vortex mixed, centrifuged to collect the aqueous phase, to obtain the primary pretreated polymyxin B.

[0060] Step 1.2: Weigh 0.375g of lysine, dissolve it in 50mL of 0.1mol / L citric acid-sodium citrate buffer solution (pH=6.6), the concentration of lysine is 0.05mM, add a certain amount of DCC and DMAP, activated at 200 rpm for 20 minutes at room temperature, wherein the mass ratio of...

Embodiment 2

[0074] The difference between this example and Example 1 is that the mass ratio of lysine to polymyxin B is 1:1.

[0075] The magnetic beads prepared in this example were used to adsorb Haemophilus influenzae type b in cerebrospinal fluid, and the adsorption efficiency was 65±2%, such as Figure 4 shown.

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Abstract

The invention discloses a polymyxin B magnetic bead, a preparation method and an application thereof. 3 o 4 Nano-magnetic beads and polymyxin B solution are mixed and then stirred at 40-60°C for 2-5 hours to obtain Fe coated with polymyxin B on the surface 3 o 4 Nano magnetic beads, that is, polymyxin B magnetic beads. The polymyxin B magnetic beads include Fe 3 o 4 Nano magnetic beads and coated in Fe 3 o 4 Polymyxin B on the surface of magnetic nanobeads. The polymyxin B magnetic beads prepared by the present invention are combined with the free negatively charged phosphate groups of lipoproteins on the cell membrane of Haemophilus influenzae type b through the specific polycationic ring of the polymyxin B molecule through electrostatic interaction, and have high accuracy. High and specific characteristics. It can quickly enrich trace amounts of Haemophilus influenzae type b from cerebral effusion and achieve good adsorption effect. At the same time, the detection efficiency and sensitivity of Haemophilus influenzae type b are improved.

Description

technical field [0001] The invention belongs to the technical field of nanomaterials and bacteria inspection, and in particular relates to a polymyxin B magnetic bead, a preparation method and an application thereof. Background technique [0002] Haemophilus influenzae type b is abbreviated as HIB, that is, haemophilus influenza type b bacteria. It is one of the main pathogenic bacteria that cause bacterial meningitis, pneumonia, bacteremia and other infectious diseases and death in young children, and seriously endangers the health of infants and young children. HIB vaccine is suitable for routine immunization of infants aged 2-17 months to prevent invasive diseases caused by HIB. Diseases caused by Haemophilus influenzae (HIB) may occur due to problems such as low absorption rate of immunization and poor vaccination effect among young children. However, there is currently a lack of effective monitoring of HIB disease and accurate assessment of disease burden in China. The...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/543G01N33/569
CPCG01N33/54326G01N33/54346G01N33/5695
Inventor 崔浪军陈蓓申萧蒙陈雪黄自华冯润东王莉芳李晓春张鹏
Owner SHAANXI NORMAL UNIV
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