An aptamer biosensor based dual signal amplification system for the detection of salmonella typhimurium

Anal Biochem. 2021 Feb 15:615:114050. doi: 10.1016/j.ab.2020.114050. Epub 2020 Dec 4.

Abstract

Salmonella, a typical foodborne pathogen, always seriously threatens the health and even life of both humans and animals. However, highly sensitive and fast quantitative methods for its detection are remaining to be challenged. Herein, we presented an efficient method with dual signal amplification strategy by combining immune hybridization chain reaction (HCR) with surface enhanced Raman scattering (SERS) to high sensitively detect Salmonella typhimurium in food. After sample preparation, S. typhimurium were specifically captured by immunomagnetic beads (IMBs), then aptamers and hairpin-probes were added to trigger HCR to form nicked dsDNA, finally 4',6-Diamidino-2-phenylindole dihydrochloride (DAPI) was incubated with HCR products and then the whole system was mixed with AgNP colloid to detect the SERS intensity at 1610 cm-1. As a result, a good linear relationship was achieved between SERS intensities and corresponding concentrations of S. typhimurium ranging from 10 to 105 CFU/mL, with a limit of detection (LOD) of 6 CFU/mL in 3.5 h. The proposed method has been successfully applied to capture and detect the S. typhimurium in spiked milk samples, and the results were consistent with those of the traditional plate counting method. The method, with combination of HCR and SERS, achieves double amplification of the detection signal and significantly improves the detection sensitivity of S. typhimurium. And it also shows good application potential for the highly sensitive detection of other contaminants in food.

Keywords: Dual signal amplification; Food safety detection; HCR; S.typhimurium; SERS.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Aptamers, Nucleotide / chemistry*
  • Biosensing Techniques / methods*
  • Food Microbiology / methods*
  • Food Safety / methods
  • Humans
  • Immunomagnetic Separation / methods
  • Limit of Detection
  • Magnetite Nanoparticles / chemistry
  • Nucleic Acid Hybridization / methods
  • Salmonella typhimurium / isolation & purification*
  • Spectrum Analysis, Raman / methods

Substances

  • Aptamers, Nucleotide
  • Magnetite Nanoparticles