We previously described isolation of a potentially new reovirus strain from the central nervous system of an 8-week-old female infant with a history of active varicella, oral thrush, hypoalbuminemia, intermittent fevers, diarrhea and feeding intolerance [Hermann et al., Ped. Inf. Dis J. 23, 373 (2004)]. This reovirus strain was tentatively identified as a member of the serotype 2 group by virus neutralization and RNA-gel electrophoresis studies and has been named type 2 Winnipeg (T2W). For this study we determined the nucleotide sequences of the T2W S1, S2, S3 and S4 genome segments to allow molecular comparison with other reoviruses. Comparative segment alignments of T2W S1 gene sequence with other reovirus S1 sequences showed T2W belongs to reovirus serotype 2. T2W S1 is most similar to the S1 genes of reovirus strains T2/Human/Netherlands/1,984 and T2/Human/Netherlands/1,973 with nucleotide identity >93%. The T2W S2 gene showed highest identity to reovirus T1 Lang S2 (approximately 75%). The T2W S3 gene showed highest identity to the S3 gene of T3/Human/Netherlands/1,983 (approximately 74%), and the T2W S4 gene showed highest identity to the T2 Jones S4 gene (approximately 73%). Pairwise protein comparisons between T2W sigma proteins and all available reovirus sigma proteins ranged from <21% identity for the sigma1 comparisons to more than 95% identity for sigma2 comparisons. The predicted T2W sigma1, sigma2 and sigma3 protein sequences were confirmed by mass spectrometry.