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Published on: July 16, 2012
Differentiation of mouse hepatitis virus genotypes by restriction fragment length polymorphism analysis
N Nakamura1, S Yoshizumi, T Urano
1Division of Microbiology and Genetics, Center for Animal Resources and Development, Institute of Resource Development and Analysis, Kumamoto University, 2-2-1 Honjo, Kumamoto 860-0811, Japan.
Abstract:
Reverse transcriptase-polymerase chain reaction (RT-PCR) amplification of the nucleocapsid (N) gene of mouse hepatitis virus (MHV) successfully detected 36 strains from the faeces of mice that had been housed in animal facilities in Japan from 2000 to 2003. Subsequent restriction fragment length polymorphism (RFLP) analysis, using the enzymes Acc I, Alu I, EcoR I and Mbo I, demonstrated that these strains could be divided into five distinct subgroups and that the same MHV strains were detected from closely related mouse facilities. Furthermore, strains from the same facility showed the same RFLP pattern, irrespective of the year of detection, but this pattern varied between different locations. This study shows that RFLP analyses are a rapid and useful method for differentiation of MHV strains.
Insights
Mouse hepatitis virus (MHV) strains were detected in mouse feces using reverse transcriptase-polymerase chain reaction (RT-PCR). Restriction fragment length polymorphism (RFLP) analysis differentiated these MHV strains into five subgroups, aiding in facility-specific tracking.
Area of Science:
- Veterinary Virology
- Molecular Epidemiology
- Animal Health Management
Background:
- Mouse hepatitis virus (MHV) is a significant pathogen affecting laboratory mice.
- Effective monitoring and differentiation of MHV strains are crucial for maintaining animal facility health and research integrity.
- Previous methods for MHV strain characterization were often time-consuming or lacked discriminatory power.
Purpose of the Study:
- To develop and validate a molecular method for differentiating MHV strains detected in Japanese animal facilities.
- To assess the utility of restriction fragment length polymorphism (RFLP) for MHV strain typing.
- To investigate the genetic diversity and distribution of MHV strains in mouse populations.
Main Methods:
- Detection of MHV using reverse transcriptase-polymerase chain reaction (RT-PCR) targeting the nucleocapsid (N) gene.
- Subtyping of detected MHV strains via restriction fragment length polymorphism (RFLP) analysis.
- Enzyme digestion with Acc I, Alu I, EcoR I, and Mbo I for RFLP profiling.
Main Results:
- Thirty-six MHV strains were successfully detected from mouse feces collected between 2000 and 2003.
- RFLP analysis delineated the detected MHV strains into five distinct subgroups.
- Consistent RFLP patterns were observed for strains from the same facility, regardless of collection year, while patterns varied between different locations.
Conclusions:
- RFLP analysis is a rapid and effective method for differentiating MHV strains.
- The findings demonstrate the potential for molecular subtyping to track MHV transmission dynamics within and between animal facilities.
- This approach aids in understanding MHV epidemiology and implementing targeted control strategies.
