Ultrasensitive Detection of Viable Enterobacter sakazakii by a Continual Cascade Nanozyme Biosensor

Anal Chem. 2017 Oct 3;89(19):10194-10200. doi: 10.1021/acs.analchem.7b01266. Epub 2017 Sep 20.

Abstract

Recent outbreaks of life-threatening neonatal infections linked to Enterobacter sakazakii (ES) heightened the need to develop rapid and ultrasensitive detection strategies, especially those capable of determining the viable cells. This study introduced a continual cascade nanozyme biosensor for the detection of viable ES based on propidium monoazide (PMA), loop-mediated isothermal amplification (LAMP), and Nanozyme strip. The ompA gene of ES was determined using FITC-modified and BIO-modified primers in the LAMP process. LAMP combined with PMA treatment was applied for distinguishing the viable from the dead state of ES. Then, using Fe3O4 magnetic nanoparticles as a nanozyme probe, a magnetic nanoparticle (MNP)-based immunochromatographic strip (Nanozyme strip) was further employed for amplifying signal to allow visual detection and quantification by a strip reader. The LAMP products were sandwiched between the anti-FITC and the anti-BIO, and the accumulation of the Fe3O4 magnetic nanoparticles enabled the visual detection of ES. The detection limit of the nanozyme biosensor was improved by 10 CFU/mL compared with previously reported techniques, and the whole manipulation process was much faster (within 1 h) and simpler (without specialist facilities). Hence, the developed continual cascade nanozyme biosensor has provided a rapid, ultrasensitive, and simple tool for on-site detection of viable ES.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Immobilized / chemistry
  • Antibodies, Immobilized / immunology
  • Azides / chemistry
  • Bacterial Outer Membrane Proteins / genetics*
  • Biosensing Techniques / methods*
  • Cronobacter sakazakii / isolation & purification
  • Cronobacter sakazakii / metabolism*
  • DNA Primers / chemistry
  • DNA Primers / metabolism
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / metabolism*
  • Ferrosoferric Oxide / chemistry
  • Fluorescein-5-isothiocyanate / chemistry
  • Immunoassay
  • Limit of Detection
  • Magnetite Nanoparticles / chemistry*
  • Nucleic Acid Amplification Techniques
  • Propidium / analogs & derivatives
  • Propidium / chemistry

Substances

  • Antibodies, Immobilized
  • Azides
  • Bacterial Outer Membrane Proteins
  • DNA Primers
  • DNA, Bacterial
  • Magnetite Nanoparticles
  • propidium monoazide
  • OMPA outer membrane proteins
  • Propidium
  • Fluorescein-5-isothiocyanate
  • Ferrosoferric Oxide