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Background   Plasmodium ovale  is relatively unfamiliar to Chinese staff engaged in malaria diagnosis. In 2013, dried blood spots of four unidentified but suspected ovale malaria samples were sent to the National Malaria Reference Laboratory (NMRL) for reconfirmation.    Methods  Partial and complete, small, subunit ribosomal DNA (SSU rDNA) sequences of four samples were obtained with PCR-cloning-sequencing method. Obtained sequences were analyzed by aligning with each other and with nine SSU rDNA sequences of six known  Plasmodium  parasites. A phylogenetic tree was constructed based on complete SSU rDNA sequences and 12 same gene sequences derived from six known  Plasmodium  parasites and three  Babesia  parasites. Primary structure of conservative and variable regions of variant sequences was determined also by comparing them with those of six known  Plasmodium  parasites. To confirm their existence in genome, they were redetected with primers matching their variable regions. PCR systems aimed to roughly detect any eukaryotes and prokaryotes respectively were also applied to search for other pathogens in one of four patients.    Results  Totally, 19 partial and 23 complete SSU rDNA sequences obtained from four samples. Except eight variant sequences, similarities among sequences from same DNA sample were in general high (more than 98%). The phylogenetic analysis revealed that three cases were infected by  P. ovale wallikeri  and one by  P. ovale curtisi . Four of the variant sequences which obtained from four samples relatively showed high similarities with each other (98.5%-100%). Identical variant sequences actually could be re-obtained from each DNA sample. Their primary structure of conservative and variable regions showed quite fit with that of six known  Plasmodium  parasites. The test for prokaryote pathogens showed negative and the tests for eukaryotes only found DNA sequences of Human and  P. ovale  parasites.    Conclusion  Both  P. ovale wallikeri  and  P. ovale curtisi  infections are present in imported malaria cases of China. New type of partial SSU rDNA sequence which assumed to express in a certain life stage of  P. ovale  was obtained from both  P. ovale wallikeri  and  P. ovale curtisi  samples. This discovery would supply information and clues to identify and understand  P. ovale  parasites more accurately.

New type of SSUrDNA sequence was detected from both Plasmodium ovale curtisi and Plasmodium ovale wallikeri samples