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Molecular identification of a recent type of canine distemper virus in Japan by restriction fragment length
K Ohashi1, K Iwatsuki, K Nakamura
1Department of Veterinary Microbiology, Graduate School of Agricultural and Sciences, University of Tokyo, Japan.
Abstract:
Restriction fragment length polymorphism analysis was used to differentiate recent field viruses of canine distemper virus (CDU) from vaccine strains. Virus genomes were amplified by using reverse transcriptase-polymerase chain reaction in part of the haemagglutinin gene. After digestion with EcoRV, the PCR products of recent field isolates were cut into two fragments that differ from the uncut form of old strains including all of vaccine strains. This method could be applied to fresh or stored brains, spleens and peripheral blood mononuclear cells of infected dogs. This molecular approach is useful for determining the causative agent of postvaccinated CDV infection.
Insights
This study developed a molecular method to distinguish canine distemper virus (CDV) field strains from vaccine strains using PCR and DNA analysis. This technique aids in identifying the cause of CDV infections after vaccination.
Area of Science:
- Veterinary Virology
- Molecular Diagnostics
- Canine Infectious Diseases
Background:
- Canine distemper virus (CDV) poses a significant threat to canine health.
- Differentiating between field and vaccine strains of CDV is crucial for accurate diagnosis and disease management.
- Current diagnostic methods may not always effectively distinguish between field and vaccine strains.
Purpose of the Study:
- To develop and validate a molecular method for differentiating recent CDV field isolates from vaccine strains.
- To provide a reliable tool for identifying the causative agent in post-vaccinated CDV infections.
Main Methods:
- Restriction fragment length polymorphism (RFLP) analysis was employed.
- Viral genomes were amplified using reverse transcriptase-polymerase chain reaction (RT-PCR) targeting the haemagglutinin gene.
- PCR products were digested with the restriction enzyme EcoRV.
Main Results:
- PCR products from recent field isolates were cleaved into two fragments by EcoRV.
- PCR products from older strains, including all vaccine strains, remained uncut.
- The method demonstrated effectiveness on various sample types like brain, spleen, and peripheral blood mononuclear cells.
Conclusions:
- The developed RFLP method provides a clear distinction between CDV field and vaccine strains.
- This molecular approach is valuable for diagnosing the specific cause of CDV infections, particularly in vaccinated dogs.
- The technique is applicable to both fresh and stored biological samples from infected dogs.