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Improving the reproducibility of the NAP1/B1/027 epidemic strain R20291 in the hamster model of infection
Michelle L Kelly1, Yen Kuan Ng1, Stephen T Cartman2
1Clostridia Research Group, BBSRC/EPSRC Synthetic Biology Research Centre (SBRC), School of Life Sciences, Centre for Biomolecular Sciences, University of Nottingham, University Park, Nottingham, NG7 2RD, United Kingdom.
Abstract:
Comparative analysis of the Clostridium difficile BI/NAP1/027 strain R20291 and ClosTron-derived ermB mutants in the hamster infection model are compromised by the clindamycin susceptibility of the parent. Mutants can appear more virulent. We have rectified this anomaly by genome engineering. The variant created (CRG20291) represents an ideal control strain for virulence assays of ClosTron mutants.
Insights
Genome engineering created a Clostridium difficile control strain (CRG20291) to accurately study ClosTron mutants. This resolves issues with clindamycin susceptibility in parent strains, enabling reliable virulence assays.
Area of Science:
- Microbiology
- Infectious Diseases
- Bacterial Genetics
Background:
- Clostridium difficile (C. diff) infection is a significant healthcare concern.
- The BI/NAP1/027 strain R20291 is a hypervirulent C. diff strain.
- ClosTron-derived ermB mutants are used to study C. diff virulence but are affected by parent strain clindamycin susceptibility.
Purpose of the Study:
- To develop a suitable control strain for C. diff virulence assays.
- To overcome limitations posed by clindamycin susceptibility in parent strains.
- To enable accurate assessment of ClosTron mutant virulence.
Main Methods:
- Comparative analysis of C. diff strain R20291 and ClosTron-derived ermB mutants.
- Utilizing genome engineering to create a modified strain.
- Employing the hamster infection model for virulence assays.
Main Results:
- The parent strain's clindamycin susceptibility compromised comparative analysis.
- ClosTron mutants initially appeared more virulent due to this anomaly.
- A genome-engineered variant (CRG20291) was successfully created.
Conclusions:
- The engineered CRG20291 strain serves as an ideal control for C. diff virulence studies.
- This resolves the anomaly caused by clindamycin susceptibility.
- Facilitates more reliable and accurate research on C. diff pathogenesis.
