title: Detection of Trypanosoma brucei parasites in blood samples using real-time nucleic acid sequence-based amplification.
abstract: Currently, the conventional diagnosis of human African trypanosomiasis (HAT) is by microscopic demonstration of trypomastigotes in blood, lymph, and/or cerebrospinal fluid. However, microscopic diagnosis of HAT is not sensitive enough and may give false-negative results, thus, denying the patient the necessary treatment of the otherwise fatal disease. For this reason, a highly sensitive technique needs to be developed to enhance case findings. In this study, the real-time nucleic acid sequence-based amplification assay described is based on amplification and concurrent detection of small subunit rRNA (18S rRNA) of Trypanosoma brucei. The sensitivity of the assay was evaluated on nucleic acid from in vitro cultured parasites and blood spiked with various parasites quantities. The assay detected 10 parasites/mL using cultured parasites as well as spiked blood. A sensitive assay such as the one developed in this study may become an alternative tool to confirm diagnosis of human African trypanosomiasis.
authors.affiliation: Faculty of Veterinary Medicine, Department of Veterinary Parasitology and Microbiology, Makerere University, Kampala, Uganda.
authors.forename: Claire M
authors.initials: CM
authors.lastname: Mugasa
authors.forename: Gerard J
authors.initials: GJ
authors.lastname: Schoone
authors.forename: Rosine A
authors.initials: RA
authors.lastname: Ekangu
authors.forename: George W
authors.initials: GW
authors.lastname: Lubega
authors.forename: Piet A
authors.initials: PA
authors.lastname: Kager
authors.forename: Henk D F H
authors.initials: HD
authors.lastname: Schallig
mesh_terms: D000818:Animals; D001769:Blood / Q000469:parasitology*; D006801:Humans; D008853:Microscopy; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D021161:Self-Sustained Sequence Replication / Q000379:methods*; D012680:Sensitivity and Specificity; D014346:Trypanosoma brucei brucei / Q000302:isolation & purification*; D014353:Trypanosomiasis, African / Q000175:diagnosis*
chemical_list: D012337:RNA, Ribosomal, 18S