Comparative genomic studies of Marek's disease virus (MDV) have suggested that attenuated and virulent strains share >
98% sequence identity. However, these estimates fail to account for variation in regions of the MDV genome harboring tandem repeats. To resolve these loci and enable assessments of intrapopulation diversity, we used a PacBio Sequel II platform to sequence MDV strains CVI988/Rispens (attenuated), HPRS-B14 (virulent), Md5 (very virulent) and 675A (very virulent plus). This approach enabled us to identify patterns of variation in tandem repeat regions that are consistent with known phenotypic differences across these strains. We also found CVI988/Rispens variants showing a 4.3-kb deletion in the Unique Short (US) region, resulting in the loss of five genes. These findings support a potential link between MDV tandem repeats and phenotypic traits like virulence and attenuation, and demonstrate that DNA viruses can harbor high levels of intrapopulation diversity in tandem repeat regions.