Detection of Acinetobacter baumannii in fresh produce using modified magnetic nanoparticles and PCR

Anal Biochem. 2020 Nov 15:609:113890. doi: 10.1016/j.ab.2020.113890. Epub 2020 Aug 9.

Abstract

Foodborne pathogens represent a major public health concern. In this respect, Acinetobacter baumannii is emerging as multi-drug resistant food pathogen. Screening of A. baumannii in food is a lengthy process. The aim of this study is to develop a fast and efficient protocol for detecting A. baumannii as well as other potential pathogens in fresh food. A. baumannii was collected from strawberry samples (n = 93) and were identified by DNA sequencing and polymerase chain reaction (PCR; recA). Hydrophobic magnetic nanoparticles (OA-MNP) were synthesized and tested for capturing A. baumannii and other bacteria from broth medium. Their ability to immobilize bacteria was assessed and the accuracy of PCR performed on immobilized bacteria was compared against control. A. baumannii (n = 14) was isolated form fresh produce as confirmed by sequencing and PCR. OA-MNPs were able to capture A. baumannii as well as other bacteria from broth medium. Intact DNA was extracted from immobilized bacteria and was successfully recruited for subsequent PCR. In conclusion, OA-MNP can be used for immobilizing food pathogens from broth medium. PCR can be performed using DNA extracted from immobilized bacteria for identification. The proposed analysis procedure may shorten the time required for detection of bacterial contamination in food.

Keywords: Acinetobacter baumannii; Antibiotic resistance; FTIR; recA gene.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acinetobacter baumannii / drug effects
  • Acinetobacter baumannii / genetics*
  • Acinetobacter baumannii / isolation & purification
  • Anti-Bacterial Agents / pharmacology
  • Bacterial Proteins / genetics
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / metabolism
  • Drug Resistance, Multiple, Bacterial / drug effects
  • Drug Resistance, Multiple, Bacterial / genetics
  • Fragaria / microbiology
  • Hydrophobic and Hydrophilic Interactions
  • Magnetite Nanoparticles / chemistry*
  • Microbial Sensitivity Tests
  • Oleic Acid / chemistry
  • Polymerase Chain Reaction / methods*
  • Sequence Analysis, DNA

Substances

  • Anti-Bacterial Agents
  • Bacterial Proteins
  • DNA, Bacterial
  • Magnetite Nanoparticles
  • Oleic Acid