title: Detection of Trypanosoma brucei spp. in human blood by a nonradioactive branched DNA-based technique.
abstract: We have developed a nonradioactive branched DNA (bDNA)-based assay for the diagnosis of the African trypanosomiases in simple buffy coat preparations of human blood. Two repetitive DNA sequences specific to the Trypanosoma brucei complex were chosen as targets of the bDNA assay, a technique which amplifies the signal from a target molecule rather than the target itself. Comparable sensitivities were observed with cloned target sequences, purified T. brucei DNA, procyclic trypanosomes, and bloodstream trypomastigotes. The results of bDNA analysis of human blood samples from Côte d'Ivoire (n = 50) showed excellent agreement with those of buffy coat microscopy. The bDNA technology offers certain advantages over alternative molecular biological techniques, including the simplicity of sample preparation and of the procedure itself, the stability of the reagents, the ability to process large numbers of samples simultaneously, and freedom from crosscontamination artifacts. We have successfully applied the bDNA technique to the detection of T. brucei in clinical samples from regions where T. brucei infection is endemic; to our knowledge, this is the first report of the molecular detection of T. brucei in human blood.
authors.affiliation: Program in Molecular Pathogenesis, University of California at San Francisco 94143-0422, USA.
authors.forename: E
authors.initials: E
authors.lastname: Harris
authors.forename: J
authors.initials: J
authors.lastname: Detmer
authors.forename: J
authors.initials: J
authors.lastname: Dungan
authors.forename: F
authors.initials: F
authors.lastname: Doua
authors.forename: T
authors.initials: T
authors.lastname: White
authors.forename: J A
authors.initials: JA
authors.lastname: Kolberg
authors.forename: M S
authors.initials: MS
authors.lastname: Urdea
authors.forename: N
authors.initials: N
authors.lastname: Agabian
mesh_terms: D000818:Animals; D001769:Blood / Q000469:parasitology*; D015342:DNA Probes; D016054:DNA, Protozoan / Q000032:analysis*; D006801:Humans; D015336:Molecular Probe Techniques; D014346:Trypanosoma brucei brucei / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D015342:DNA Probes; D016054:DNA, Protozoan