Analysis of the entire genomes of thirteen TT virus variants classifiable into the fourth and fifth genetic groups, isolated from viremic infants

TT virus (TTV) DNA in serum samples obtained from 24 TTV-infected infants was amplified by polymerase chain reaction (PCR) with inverse primers derived from the untranslated region. The amplified PCR products were molecularly cloned; six clones each were analyzed. Seventy-six (53%) of the 144 TTV clones were classified into group 4 (YONBAN isolates), and 22 (15%) into a novel genetic group (group 5). The TTV clones in group 4 were classified into 9 types, and those in group 5 into 4 types. The entire nucleotide sequence of one representative clone each from the 13 types were determined; they comprised 3570-3770 nucleotides, and had poor homology to TTVs of groups 1-3 (TA278, PMV and SANBAN isolates). A phylogenetic tree based on the entire nucleotide sequence of open reading frame 1 confirmed the presence of five distinct clusters separated by a bootstrap value of 100%. Analysis of 13 TTV variants demonstrated preservation of the genomic organization and transcription profile in all TTV groups. TTV group 4 was detected in 54% or 72% of 7-to-12-month-old infants in Japan and China, respectively, which is comparable with that among adults in the respective country, indicating early and frequent acquisition of this TTV group in infancy.