Streptavidin-exposed magnetic nanoparticles for lectin magnetic separation (LMS) of Staphylococcus aureus prior to three quantification strategies

Mikrochim Acta. 2019 Nov 19;186(12):813. doi: 10.1007/s00604-019-3978-4.

Abstract

A lectin magnetic separation (LMS) method for Staphylococcus aureus (S. aureus) was developed with the aim to improve the efficiency of magnetic nanoparticles and to expand the scope of bacterial recognition. Poly(ethylene glycol) (PEG)-mediated magnetic nanoparticles modified with streptavidin (MNP-PEG-SA) were synthesized and then applied to a two-step LMS based on the use of wheat germ agglutinin (WGA). Three specific methods for S. aureus detection (suitable for different requirements including detection time and sensitivity) were designed. The new LMS has improved anchoring efficiency (compared to two-step LMS methods) and requires a reduced number of magnetic particles. The Baird-Parker (B-P) method can detect S. aureus with a detection limit of 3 × 100 CFU·mL-1 within 15 h; the polymerase chain reaction (PCR) method can be finished within 4 h, with the lowest detection limit (LOD) of 3 × 102 CFU·mL-1. The LOD of HRP-pig IgG-based colorimetric method is 3 × 105 CFU·mL-1, and the method only lasts for 2 h. If combined with specific detection methods, it meets different needs for rapid detection of S. aureus. Graphical abstractSchematic representation of lectin magnetic separation (LMS) based on biotin-wheat germ agglutinin (WGA) and poly (ethylene glycol) (PEG)-mediated streptavidin-modified magnetic nanoparticles (MNP-PEG-SA) and three different quantification strategies (including B-P culture assay, PCR assay, and colorimetric assay) for S. aureus.

Keywords: Baird–Parker culture; HRP-pig IgG-based colorimetric reaction; Lectin magnetic separation (LMS); Polymerase chain reaction; Staphylococcus aureus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Armoracia / enzymology
  • Benzidines / chemistry
  • Biotin / chemistry
  • Colorimetry / methods
  • Food Contamination / analysis
  • Fruit and Vegetable Juices / microbiology
  • Horseradish Peroxidase / chemistry
  • Hydrogen Peroxide
  • Immunoglobulin G / chemistry
  • Immunomagnetic Separation / methods
  • Limit of Detection
  • Magnetic Phenomena
  • Nanoparticles / chemistry*
  • Polymerase Chain Reaction / methods
  • Staphylococcus aureus / isolation & purification*
  • Streptavidin / chemistry*
  • Swine
  • Triticum / chemistry
  • Wheat Germ Agglutinins / chemistry*

Substances

  • Benzidines
  • IGG-horseradish peroxidase
  • Immunoglobulin G
  • Wheat Germ Agglutinins
  • 3,3',5,5'-tetramethylbenzidine
  • Biotin
  • Streptavidin
  • Hydrogen Peroxide
  • Horseradish Peroxidase