Objective To study tissue tropism of TT virus in experimentally infected Rhesus monkey. Methods Various tissues were collected from 5 experimentally infected monkeys during the viremic period. Total DNA was extracted from tissues of 5 experimentally infected Rhesus monkeys. A dot hybridization was done with virus double DNA strand probe or single antisense strand probe. Results The double strand probe was hybridized with DNA of liver, bone marrow, spleen, stomach, small intestine and colon. The single strand antisense probe was hybridized only with DNA of liver, small intestine and bone marrow of all 5 monkeys, but not with that of other tissues.Conclusions As the viral genome is of negative polarity, the plus stranded fragment identified in our study might be a replicative intermediate, and is only demonstrated in liver, small intestine, and bone marrow by dot blot hybridization with single stranded antisense probe. It suggests that TT virus might be, tropism of liver, small intestine, and bone marrow, and replicate in tissue mentioned.