Isolation of Campylobacter From Human Stool Samples

Indian J Med Microbiol. Jan-Mar 2014;32(1):35-8. doi: 10.4103/0255-0857.124294.

Abstract

Context: Campylobacter is an undetected cause of diarrhoea especially under 5 years of age in most of the countries. Isolation of this organism is difficult, expensive and cumbersome.

Aims: Our objective of this study was to isolate this pathogen from the stool specimens on routinely available blood containing laboratory media using the candle jar for creating the microaerophilic atmosphere in our setup.

Settings and designs: A descriptive study.

Materials and methods: A total of 50 stool samples were inoculated onto selective and non-selective media with and without filtration using a 0.45 μm membrane. The inoculated media were simultaneously incubated in microaerophilic conditions using the Anoxomat as well as in candle jars at temperatures 37°C and 42°C. The culture isolates were confirmed by standard phenotypic tests. A simplex polymerase chain reaction (PCR) targeting the 16S ribosomal deoxyribonucleic acid of Campylobacter was performed on the deoxyribonucleic acid (DNA) of the culture isolates as well as on the DNA extracted from the stool filtrates.

Statistical analysis: Data was expressed as a proportion.

Results: Campylobacter could be isolated in 5 out of 50 stool samples using both the Anoxomat as well as the candle jar. Furthermore, we did not find any difference between the isolation using the selective and blood containing media as well as the different incubation temperatures. All the five were confirmed phenotypically and genotypically to be Campylobacter jejuni. The PCR results corroborated with that of the culture.

Conclusions: Isolation by culture was as sensitive as that of the PCR.

Publication types

  • Comparative Study
  • Evaluation Study

MeSH terms

  • Adolescent
  • Bacteriological Techniques / methods*
  • Campylobacter Infections / diagnosis*
  • Campylobacter jejuni / isolation & purification*
  • Child
  • Child, Preschool
  • Culture Media
  • Feces / microbiology*
  • Female
  • Humans
  • Infant
  • Infant, Newborn
  • Male
  • Molecular Diagnostic Techniques / methods*
  • Polymerase Chain Reaction

Substances

  • Culture Media