Magnetic microsphere and preparation method thereof, and microorganism detection method

A magnetic microsphere and detection method technology, applied in the field of microbial detection, can solve the problems of expensive technology, low sensitivity, time-consuming, etc., and achieve the effect of reducing interference and saving bacterial enrichment

Pending Publication Date: 2019-05-21
ACADEMY OF MILITARY MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As a supplement to traditional detection methods in bacterial identification and drug susceptibility testing of pathogenic bacteria, there are still many problems that limit its use
False negatives or false positives that occur during PCR identification and the multi-step chemistry required during immunoassays limit their applications, which are expensive, time-consuming and / or exhibit low sensitivity

Method used

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  • Magnetic microsphere and preparation method thereof, and microorganism detection method
  • Magnetic microsphere and preparation method thereof, and microorganism detection method
  • Magnetic microsphere and preparation method thereof, and microorganism detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] 1. Chemicals and materials

[0078] Polyethyleneimine branched (PEI, MW 25kDa), silver nitrate (AgNO 3 ) were obtained from Sigma-Aldrich. Buy iron(III) chloride hexahydrate (FeCl) from Sinopharm Chemical Reagent Co. 3 ·6H 2 O), polyethylene glycol (PEG6000), trisodium citrate (TSC), ethylene glycol (EG) and anhydrous sodium acetate (NaAc). Other reagents were from Shanghai Chemical Reagent Co., Ltd. (China). All chemicals are of analytical grade.

[0079] Aqueous solutions were prepared with Millipore ultrapure water and purified in a Milli-Q system (18.2 MΩcm-1). Clinical isolates of Staphylococcus aureus, Pseudomonas aeruginosa and Acinetobacter baumannii were collected from the Affiliated Hospital of Xuzhou Medical University. Methicillin (S. aureus) and imipenem (Pseudomonas aeruginosa, Acinetobacter baumannii) resistance was defined according to the Clinical and Laboratory Standards Institute breakpoint criteria (CLSI M100, S27). All participants provided i...

Embodiment 2

[0098] 1. Antibiotic sensitivity test

[0099] Detection of susceptibility or resistance to a drug or drug class by agar dilution test, primarily antimicrobial susceptibility testing using Mueller-Hinton agar (MHA). Prepare a 10× concentration of antimicrobial at 1280 μg / mL and serially dilute to twice the desired final concentration. To prepare the intermediate diluent, a convenient formula to use is C1*V1=C2*V2, where C1 is the concentration of the antimicrobial stock solution (typically 1280 µg / mL or higher) and V1 is the unknown required to produce the intermediate concentration Volume, C2 is the intermediate concentration, and V2 is the volume of the intermediate stock solution we need. The solvent and diluent of imipenem is phosphate buffer (pH7.2, 0.01mol / L). The solvent and diluent for oxacillin is water. Dilute 1:10 with Mueller-Hinton agar to obtain the final concentration for antibiotic susceptibility agar plates. MRSA was detected using MHA containing 4% NaCl a...

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Abstract

The invention relates to the field of microorganism detection, in particular to a magnetic microsphere and a preparation method thereof, and a microorganism detection method. The magnetic microspherecomprises a microsphere body which carries negative charges, and a PEI adsorbed on the outer surface of the microsphere body and carrying positive charges. The magnetic microsphere provided by the invention can effectively adsorb microorganisms in a sample; the method has the advantages that the specificity is strong, the adsorption is tight, the interference of other impurities is reduced, different microorganisms are distinguished by detecting Raman signals, the detection time is greatly shortened, the method is convenient and reliable, the accuracy rate is 96% or above, and a novel and rapid detection mode is provided for the diagnosis and treatment of clinical blood infection.

Description

technical field [0001] The invention relates to the field of microorganism detection, in particular to a magnetic microsphere, a preparation method thereof and a microorganism detection method. Background technique [0002] In recent years, with the vigorous development of invasive diagnosis and treatment techniques and the wide application of broad-spectrum antibiotics and hormones, the incidence of bloodstream infection has increased year by year. The mortality rate of bloodstream infection is high, the morbidity rate is high, the hospitalization time is long, the medical expenses are high, and the harm is serious. Therefore, the control of bloodstream infections has received increasing attention. With the development of various operating techniques and the application of antibacterial drugs, the pathogens causing bloodstream infections are constantly changing, and the drug resistance of pathogens is gradually increasing. Methicillin-resistant Staphylococcus aureus (MRSA...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/65
Inventor 肖瑞汪崇文李佳
Owner ACADEMY OF MILITARY MEDICAL SCI
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