In the present investigation we molecularly characterized nontypeable rotavirus strains previously identified during surveillance in New Delhi, India. The majority of strains were demonstrated to belong to genotype G1 (54.5%) or P[8] (77.8%) on the basis of nucleotide sequencing of fragments from their VP7 and VP4 genes. The other genotypes detected included G2, G8, G9, G12, and P[4]. A G8P[6] strain, strain DS108, was detected for the first time in northern India. The VP7 gene of DS108 was most homologous with the VP7 gene of a bovine G8 strain, strain A5 (98.9%), indicating its bovine parentage. In contrast, the VP4 gene had a high degree of nucleotide sequence homology (92.9% to 99.1%) with the VP4 genes of human P[6] strains. The VP6 gene and nonstructural genes (NSP1 to NSP3 and NSP5) were most homologous with the VP6 gene and nonstructural genes of human rotaviruses belonging to the DS1 genogroup. Interestingly, the NSP4 gene of DS108 clustered within genotype E6 that until now had only two representative strains, both with G12P[6] specificity (strains RV176-00 and N26-02). Together, these results indicate that G8 strain DS108 belongs to the DS1 genogroup and could be the result of the acquisition of the VP7, VP4, and NSP4 genes by a human G2P[4] strain from more than one donor, similar to the evolution of G12P[6] strain RV176-00. The present study highlights the importance of characterizing multiple genes of nontypeable rotavirus strains to detect novel strains and get a more complete picture of rotavirus evolution.Rotaviruses (RVs) are the most important etiological agents of severe dehydrating diarrhea in children, claiming approximately 611,000 lives each year (27). Group A RVs belong to the family Reoviridae and possess a genome of 11 segments of double-stranded RNA. The two major surface proteins, the VP7 (G type) glycoprotein and protease-cleaved protein VP4 (P type), form the basis of a dual classification system for RV. To date, 23 G types and 31 P types have been identified (35,39). Worldwide surveillance studies have reported on the continued predominance of G1 to G4 and G9 RV strains (2, 34). Additionally, G12 RVs were recently reported in neonatal and diarrheal children at high prevalence rates on the Indian subcontinent and may be emerging globally (30,32,36,38). Some RV genotypes may be common in particular regions or countries and may be detected only sporadically elsewhere; for example, G5 is predominant in Brazil and G8 is predominant on the African continent (5, 7, 10, 22).RV infection in India accounts for both high rates of mortality and high rates of morbidity, leading to approximately 400,000 hospitalizations each year (2, 15). A study recently conducted by Mendelsohn et al. observed that RV is the cause of considerable economic burden in India and suggested that an appropriate effective RV vaccine may provide significant economic savings (24). Two live oral vaccines-Rotateq, a human-bovine pentavalent vaccine, and Rotarix, a G1P[8] monovalent human rotavirus vaccine-have already bee...