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December 30, 2025Journal of Veterinary Medical ScienceOpen Access

High-throughput detection of bovine leukocyte antigen (BoLA)-DRB3 alleles associated with resistance to bovine leukemia virus using a rapid allele-specific PCR assay

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Authors

RNRin NAGAIYOYao OUTOTomoyuki Onda

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Overview

Diagnostic validation study demonstrates high-accuracy detection of leukemia-resistant BoLA-DRB3 alleles in cattle, suggesting a practical tool for genetic herd screening.

Key Points

  • To develop and validate a rapid, cost-effective allele-specific PCR assay to detect BoLA-DRB3 alleles associated with bovine leukemia virus resistance in cattle.
  • Designed an allele-specific PCR assay using sequence-specific primers targeting DRB3*009:02, DRB3*014:01:01, and DRB3*002:01 alleles, validated against PCR-sequence-based typing.
  • Evaluated sensitivity and specificity using synthesized and genomic DNA, and screened field samples from N=444 cattle using a two-stage pooling approach and standard agarose gel electrophoresis.
  • The assay demonstrated 100% sensitivity and 94% specificity compared with PCR-sequence-based typing, with a detection limit of 0.8 ng/reaction.
  • High-throughput screening of 444 cattle revealed that 27.0% carried at least one resistance-associated target allele.

Cite This Study

NAGAI et al. (2025) studied this question.

synapsesocial.com/papers/6aaf66a9debf0b2b005d2c53https://doi.org/10.1292/jvms.25-0429
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