Validation and improvement of a multiplex PCR method to detect murine Helicobacter species in feces samples of mice.
multiplex PCR
non H. pylori Helicobacter
validity
Journal
Helicobacter
ISSN: 1523-5378
Titre abrégé: Helicobacter
Pays: England
ID NLM: 9605411
Informations de publication
Date de publication:
Jun 2022
Jun 2022
Historique:
revised:
28
02
2022
received:
01
09
2021
accepted:
15
03
2022
pubmed:
2
4
2022
medline:
14
5
2022
entrez:
1
4
2022
Statut:
ppublish
Résumé
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. 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. The method established was highly sensitive and specific. All five strains were detectable with a detection limit of 10 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.
Sections du résumé
BACKGROUND
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
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
RESULTS
The method established was highly sensitive and specific. All five strains were detectable with a detection limit of 10
CONCLUSION
CONCLUSIONS
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.
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
e12888Subventions
Organisme : German Center for Infection Research
Informations de copyright
© 2022 John Wiley & Sons Ltd.
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