Rapid identification of Salmonella using Hektoen enteric agar and 16s ribosomal DNA probe-gold nanoparticle immunochromatography assay in clinical faecal specimens

Lett Appl Microbiol. 2014 Apr;58(4):311-7. doi: 10.1111/lam.12191. Epub 2013 Nov 29.

Abstract

A rapid identification of Salmonella, one of the most common foodborne pathogens worldwide, in clinical patients can enable better rational managements and prevent further outbreaks. The traditional immunochromatography using antibody-gold nanoparticles (Ab-AuNPs), such as the home pregnancy test, has been used for the Salmonella detection. In this study, we developed a new and rapid method using DNA probe-AuNPs for the detection of 16s ribosomal DNA of Salmonella. To evaluate the proposed method in clinical specimens, we performed a clinical test by identifying 159 stool samples on Hektoen agar containing black or crystalloid colonies using the method and the VITEK 2 system for confirmation. Eighty of the isolates were correctly identified as Salmonella to achieve 100% sensitivity. Seventy-five samples were correctly identified as non-Salmonella spp., but four were incorrectly identified as Salmonella. The specificity was 94·93%. The assay time is about 30 min after the DNA purification. The time-consuming and labour-intense biochemical tests can be replaced. We demonstrated that this assay is a rapid, convenient and cost-effective tool for Salmonella identification of clinical faecal samples, which is worth for further promotion and clinical use. This is the first application of using 16s ribosomal DNA probe-Au-NPs and immunochromatography on clinical samples.

Significance and impact of the study: This is the first application of using 16s ribosomal DNA probe-gold nanoparticles and immunochromatography method on clinical samples with sensitivity 100% and specificity 94·93%. The assay time is about 30 min after the DNA purification. We find this assay a rapid, convenient, sensitive and inexpensive tool for Salmonella identification of clinical faecal samples, which is worth further promotion and clinical use and can replace the traditional time-consuming and labour-intense biochemical tests. The potential benefit of this approach is to develop a rapid point-of-care test that provides results while the patient is still at the doctors' office.

Keywords: Salmonella; gold nanoparticles; immunochromatography; lateral flow assay; molecular identification.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Agar
  • Base Sequence
  • Chromatography, Affinity / methods*
  • DNA Probes
  • DNA, Ribosomal
  • Feces / microbiology*
  • Gold / chemistry
  • Humans
  • Metal Nanoparticles
  • Molecular Sequence Data
  • RNA, Ribosomal, 16S / genetics
  • Salmonella / classification
  • Salmonella / genetics
  • Salmonella / isolation & purification*
  • Sensitivity and Specificity

Substances

  • DNA Probes
  • DNA, Ribosomal
  • RNA, Ribosomal, 16S
  • Gold
  • Agar

Associated data

  • GENBANK/Z49264