Ultrafast and Ultrasensitive Naked-Eye Detection of Urease-Positive Bacteria with Plasmonic Nanosensors

ACS Sens. 2019 Apr 26;4(4):961-967. doi: 10.1021/acssensors.9b00063. Epub 2019 Mar 22.

Abstract

Identifying the pathogen responsible for an infection is a requirement in order to personalize antimicrobial treatments. Detecting bacterial enzymes, such as proteases, lipases, and oxidoreductases, is a winning approach for detecting pathogens at the point of care. In this Article, a new method for detecting urease-producing bacteria rapidly and at ultralow concentrations is reported. In this method, longsome bacteriological culture steps are substituted for a 10 min capture procedure with positively charged magnetic beads. The presence of urease-positive bacteria on the particles is then queried with a plasmonic signal generation step that generates blue- or red-colored nanoparticle suspensions upon addition of the enzyme substrate. These colorimetric signals, which can be easily identified by eye, are generated by the NH3-dependent assembly of gold nanoparticles in the presence of bovine serum albumin (BSA). The proposed method can detect Proteus mirabilis with a limit of detection of 101 cells mL-1, with a total assay time of 40 min, even in the presence of a large excess of urease-negative bacteria ( Pseudomonas aeruginosa). Furthermore, it does not require bulky equipment, and it can detect P. mirabilis at clinically relevant concentrations within minutes, making it suitable for detecting urease-positive pathogens at the point of care.

Keywords: LSPR; colorimetric; gold; magnetic; nanoparticle; pathogen; sepsis; urine.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Ammonia / chemistry
  • Animals
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / urine*
  • Bacterial Typing Techniques / methods*
  • Cattle
  • Colorimetry / methods
  • Enzyme Assays / methods
  • Gold / chemistry
  • Limit of Detection
  • Magnetic Phenomena
  • Metal Nanoparticles / chemistry*
  • Polyethylenes / chemistry
  • Proteus mirabilis / enzymology
  • Proteus mirabilis / isolation & purification*
  • Quaternary Ammonium Compounds / chemistry
  • Serum Albumin, Bovine / chemistry
  • Surface Plasmon Resonance / methods
  • Urea / chemistry
  • Urease / chemistry
  • Urease / urine*
  • Urine / microbiology

Substances

  • Bacterial Proteins
  • Polyethylenes
  • Quaternary Ammonium Compounds
  • poly-N,N-dimethyl-N,N-diallylammonium chloride
  • Serum Albumin, Bovine
  • Gold
  • Ammonia
  • Urea
  • Urease