In order to better understand phylogenetic relationships among tintinnid ciliated protozoa, we sequenced and analyzed the SSU rDNA and ITS1-5.8S-ITS2 regions of 10 species belonging to five genera in the order Tintinnida. The secondary structures of the ITS2 region were compared among 8 closely related genera, revealing two stable helices of the palm. In addition, we identified a bulge absence in position II of the ITS2 putative secondary structures of species in basal positions in phylogenetic trees, suggesting the absence bulge might be an ancestral character in the order Tintinnida. Phylogenetic analyses based on SSU rDNA and ITS1-5.8S-ITS2 regions sequence show 1) divergences within the family Tintinnidae are higher than that among other four families (Codonellidae, Ptychocylididae, Metacylididae and Codonellopsidae), suggesting the subdivision of the this family; 2) the family Ptychocylididae is polyphyletic; 3) the subdivision of Tintinnopsis are suggested, because the Tintinnopsis spp. scatter into different clades; 4) species with agglutinated loricae are not clearly separated from that with hyaline ones.