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Resolution of complex feline leukocyte antigen DRB loci by reference strand-mediated conformational analysis (RSCA).
L J Kennedy1, R Ryvar, J J Brown
1Centre for Integrated Genomic Medical Research, University of Manchester, Manchester, UK. Lorna.Kennedy@man.ac.uk
Tissue Antigens
|September 17, 2003
Summary
Reference strand-mediated conformational analysis (RSCA) offers a reproducible method to study feline FLA-DRB gene polymorphism. This technique identified more alleles than previous methods, aiding research into feline health and disease resistance.
Area of Science:
- Immunogenetics
- Veterinary Genetics
Background:
- Feline leukocyte antigen (FLA)-DRB genes are highly polymorphic, with previous studies identifying 24 alleles using complex clonal sequence analysis.
- The complexity and susceptibility to artifacts in clonal sequence analysis have hindered further research into FLA-DRB genetic diversity.
Purpose of the Study:
- To apply and evaluate reference strand-mediated conformational analysis (RSCA) for characterizing FLA-DRB genetic polymorphism in domestic cats.
- To compare the efficacy of RSCA with clonal sequence analysis in identifying FLA-DRB alleles and assessing genetic variation.
Main Methods:
- Reference strand-mediated conformational analysis (RSCA) was employed using five reference strands, including those from non-domestic felines.
- Genetic polymorphism of FLA-DRB was investigated in 71 domestic cats, with 31 of these also analyzed using clonal sequence analysis.
- Allele patterns were analyzed to identify breed-specific distributions and variations in gene copy number within families.
Main Results:
- RSCA successfully distinguished 23 FLA-DRB alleles and proved highly reproducible.
- On average, RSCA identified 0.9 more alleles per cat compared to clonal sequence analysis.
- Significant differences in FLA-DRB allele distributions were observed between cat breeds, with the Burmese breed showing potentially restricted polymorphism. Evidence for variable DRB gene numbers (two or three) on different haplotypes was found.
Conclusions:
- RSCA is a valuable, rapid, and reproducible method for resolving complex amplicons and characterizing FLA-DRB genetic diversity in cats.
- This technique facilitates the identification of novel alleles and enables comprehensive studies on feline vaccine response and susceptibility to viral infections.