16S rRNA PCR followed by restriction endonuclease digestion: a rapid approach for genus level identification of important enteric bacterial pathogens

J Microbiol Methods. 2013 Dec;95(3):353-6. doi: 10.1016/j.mimet.2013.10.001. Epub 2013 Oct 16.

Abstract

The study describes a rapid approach for detection of common enteric bacterial pathogens, which involves partial amplification of the 16S rRNA gene by PCR using a colony from selective medium followed by restriction enzyme (RE) digestion using the EcoRI, HindIII and SalI enzymes. On the basis of RE digestion analysis different genera namely, Escherichia, Salmonella, Shigella, Vibrio, Campylobacter, Arcobacter, Yesinia and Listeria were differentiated.

Keywords: 16S rRNA; Diarrhea; Enteric bacterial pathogen; RE digestion.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteria / classification
  • Bacteria / genetics
  • Bacteria / isolation & purification*
  • Bacteriological Techniques / methods*
  • DNA Restriction Enzymes / genetics
  • DNA Restriction Enzymes / metabolism
  • DNA, Bacterial / genetics
  • DNA, Bacterial / metabolism
  • Gastrointestinal Diseases / diagnosis*
  • Gastrointestinal Diseases / microbiology
  • Gram-Negative Bacterial Infections / diagnosis
  • Gram-Negative Bacterial Infections / microbiology
  • Gram-Positive Bacterial Infections / diagnosis
  • Gram-Positive Bacterial Infections / microbiology
  • Molecular Diagnostic Techniques / methods*
  • Polymerase Chain Reaction / methods*
  • Polymorphism, Restriction Fragment Length*
  • RNA, Ribosomal, 16S / genetics

Substances

  • DNA, Bacterial
  • RNA, Ribosomal, 16S
  • DNA Restriction Enzymes