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Updated: Aug 18, 2026
Mouse Models Of Helicobacter Infection And Gastric Pathologies
Published on: October 18, 2018
Differential detection of five mouse-infecting helicobacter species by multiplex PCR
Sunlian Feng1, Karin Ku, Emir Hodzic
1Center for Comparative Medicine, School of Medicine, University of California, Davis, Davis, California 95616, USA.
Abstract:
Several species of helicobacter have been isolated from laboratory mice, including H. bilis, H. hepaticus, H. muridarum, H. rodentium, and H. typhlonius, which appear to be the most common. The most widely used published method for molecular detection of these agents is PCR amplification of a conserved region of 16S rRNA, but differential speciation requires restriction enzyme digestion of the amplicons. This study was undertaken to determine PCR conditions that would simultaneously and specifically identify each of the five common species without restriction enzyme analyses. First, we designed novel and specific PCR primers for H. bilis, H. hepaticus, H. muridarum, H. rodentium, and H. typhlonius, using sequences from the heterologous regions of 16S rRNA. Because of comigration of amplified products, we next identified P17, an H. bilis-specific protein; P25, an H. hepaticus-specific protein; and P30, an H. muridarum-specific protein by screening genomic DNA expression libraries of each species. Primers were designed from these three genes, plus newly designed, species-specific 16S rRNA primers for H. rodentium and H. typhlonius that could be utilized for a five-plex PCR. The sizes of the amplicons from H. bilis, H. hepaticus, H. muridarum, H. rodentium, and H. typhlonius were 435, 705, 807, 191, and 122 bp, respectively, allowing simultaneous detection and effective discrimination among species.
Insights
This study developed a novel multiplex PCR method for simultaneously identifying five common Helicobacter species in mice. This technique eliminates the need for restriction enzyme digestion, improving diagnostic efficiency for H. bilis, H. hepaticus, H. muridarum, H. rodentium, and H. typhlonius.
Area of Science:
- Veterinary Microbiology
- Molecular Diagnostics
- Mouse Health
Background:
- Several Helicobacter species (H. bilis, H. hepaticus, H. muridarum, H. rodentium, H. typhlonius) are common in laboratory mice.
- Current molecular detection relies on 16S rRNA PCR followed by restriction enzyme digestion for species differentiation.
- This method is laborious and time-consuming, necessitating a more streamlined approach.
Purpose of the Study:
- To develop a multiplex PCR assay for the simultaneous and specific detection of five common Helicobacter species in mice.
- To eliminate the requirement for post-PCR restriction enzyme analysis.
- To improve the efficiency and accuracy of Helicobacter species identification in laboratory animal diagnostics.
Main Methods:
- Designed novel, species-specific PCR primers targeting unique regions of 16S rRNA genes for H. rodentium and H. typhlonius.
- Identified species-specific protein markers (P17 for H. bilis, P25 for H. hepaticus, P30 for H. muridarum) and designed primers based on these genes.
- Developed a five-plex PCR assay combining these novel primers for simultaneous amplification and detection.
Main Results:
- Successfully developed species-specific primers for all five target Helicobacter species.
- Achieved simultaneous amplification in a single multiplex PCR reaction.
- Generated distinct amplicon sizes (435 bp for H. bilis, 705 bp for H. hepaticus, 807 bp for H. muridarum, 191 bp for H. rodentium, 122 bp for H. typhlonius) allowing clear differentiation.
Conclusions:
- The developed five-plex PCR assay enables rapid, simultaneous, and specific identification of five common Helicobacter species in mice.
- This method obviates the need for restriction enzyme digestion, significantly enhancing diagnostic workflow.
- This assay provides a valuable tool for monitoring and controlling Helicobacter infections in research mouse colonies.
