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Updated: Sep 28, 2025
Mouse Models Of Helicobacter Infection And Gastric Pathologies
Published on: October 18, 2018
Validation and improvement of a multiplex PCR method to detect murine Helicobacter species in feces samples of mice
Vanessa Neubert1, Ahmed Sadek1, Teresa Burell1
1Institute for Medical Microbiology, Immunology and Hygiene, Technical University of Munich, Munich, Germany.
Background:
Murine Helicobacter species have gained increasing awareness in mouse facilities over the last years. Infections with Helicobacter species may have an impact effect on the health of mice and might pose a zoonotic risk to researchers. To minimize the interference with experiments and hence contribute to the 3Rs, a reliable method of monitoring Helicobacter infections in animal facilities needs to be available. The aim of this study was to improve and validate the detection of the most common murine Helicobacter species.
Material And Methods:
A multiplex PCR assay was developed for identification of Helicobacter hepaticus, H. bilis, H. muridarum, H. rodentium, and H. typhlonius that could simultaneously detect these five strains in fecal samples. To ensure the quality of the results, the method was validated based on recommendations for in-house developed tests.
Results:
The method established was highly sensitive and specific. All five strains were detectable with a detection limit of 102 bacteria. Eight different mouse facilities were tested with the validated assay, and the following prevalence were found: H. rodentium 57%, H. hepaticus 46%, H. typhlonius 17%, H. bilis 12%, and H. muridarum 0%.
Conclusion:
The multiplex PCR is a reliable, economic, and time-saving diagnostic tool for routine health monitoring. Further prevalence studies are needed to confirm the high prevalence and hence importance of H. rodentium, as until now this agent is not yet listed in FELASA recommendations.
Insights
A new multiplex PCR assay reliably detects common Helicobacter species in mice, crucial for animal health monitoring and reducing experimental interference. This method aids in understanding infection prevalence and ensuring laboratory safety.
Area of Science:
- Veterinary Microbiology
- Animal Health Monitoring
- Zoonotic Disease Research
Background:
- Murine Helicobacter species infections are increasingly recognized in mouse facilities.
- These infections impact mouse health and pose potential zoonotic risks to researchers.
- Accurate monitoring is essential for experimental integrity and the 3Rs (Replacement, Reduction, Refinement).
Purpose of the Study:
- To develop and validate an improved detection method for common murine Helicobacter species.
- To establish a reliable diagnostic tool for routine health monitoring in animal facilities.
Main Methods:
- A multiplex PCR assay was designed to simultaneously identify five key Helicobacter species (H. hepaticus, H. bilis, H. muridarum, H. rodentium, H. typhlonius).
- Fecal samples were analyzed using the developed assay.
- The method underwent validation according to in-house test development guidelines.
Main Results:
- The multiplex PCR assay demonstrated high sensitivity and specificity, detecting all five target strains.
- The detection limit was as low as 10^2 bacteria per sample.
- Prevalence in eight tested mouse facilities: H. rodentium (57%), H. hepaticus (46%), H. typhlonius (17%), H. bilis (12%), H. muridarum (0%).
Conclusions:
- The validated multiplex PCR assay is a dependable, cost-effective, and time-efficient tool for routine health surveillance.
- Findings highlight the high prevalence of H. rodentium, suggesting a need for further studies and potential inclusion in FELASA recommendations.
- The assay contributes to minimizing experimental interference and enhancing animal welfare.
