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Author Spotlight: Development of an Enhanced Protocol for Rapid and Accurate Isolation of Campylobacter from Food Products
Published on: February 23, 2024
Isolation of Campylobacter from human stool samples
S M Salim, J Mandal1, S C Parija
1Department of Microbiology, Jawaharlal Institute of Postgraduate Medical Education and Research, Puducherry, India.
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.
Insights
Campylobacter jejuni, a common cause of diarrhea, can be effectively isolated from stool samples using standard laboratory media and a candle jar. This method proved as sensitive as polymerase chain reaction (PCR) for detection.
Area of Science:
- Microbiology
- Clinical Diagnostics
- Infectious Diseases
Background:
- Campylobacter is a significant, often undetected, cause of diarrhea in children under five.
- Standard isolation methods for Campylobacter are difficult, expensive, and time-consuming.
Purpose of the Study:
- To evaluate the efficacy of isolating Campylobacter from stool specimens using routine blood-containing media and a candle jar for microaerophilic atmosphere generation.
- To compare this method with standard laboratory techniques and polymerase chain reaction (PCR).
Main Methods:
- A descriptive study involving 50 stool samples.
- Inoculation onto selective and non-selective media, with and without 0.45 μm membrane filtration.
- Incubation under microaerophilic conditions using Anoxomat and candle jars at 37°C and 42°C.
- Confirmation by phenotypic tests and simplex PCR targeting 16S ribosomal DNA.
Main Results:
- Campylobacter was isolated from 5 out of 50 stool samples using both Anoxomat and candle jar methods.
- No significant difference was observed between selective and non-selective media or incubation temperatures.
- All isolates were confirmed as Campylobacter jejuni phenotypically and genotypically.
- PCR results correlated with culture findings.
Conclusions:
- Culture isolation of Campylobacter using routine media and a candle jar is a viable and sensitive method.
- This approach is comparable in sensitivity to PCR for Campylobacter detection.
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