Documenten

  • 2014jmdi0400

    !!Final published version, 538 KB, PDF-document

DOI

Trypanosomatids transcribe their genes in large polycistronic clusters that are further processed into mature mRNA molecules by trans-splicing. During this maturation process, a conserved spliced leader RNA (SL-RNA) sequence of 39 bp is physically linked to the 5' end of the pre-mRNA molecules. Trypanosomatid infections cause a series of devastating diseases in man (sleeping sickness, leishmaniasis, Chagas disease) and animals (nagana, surra, dourine). Here, we investigated the SL-RNA molecule for its diagnostic potential using reverse transcription followed by real-time PCR. As a model, we used Trypanosoma brucei gambiense, which causes sleeping sickness in west and central Africa. We showed that the copy number of the SL-RNA molecule in one single parasitic cell is at least 8600. We observed a lower detection limit of the SL-RNA assay in spiked blood samples of 100 trypanosomes per milliliter of blood. We also proved that we can detect the trypanosome's SL-RNA in the blood of sleeping sickness patients with a sensitivity of 92% (95% CI, 78%-97%) and a specificity of 96% (95% CI, 86%-99%). The SL-RNA is thus an attractive new molecular target for next-generation diagnostics in diseases caused by trypanosomatids.

Originele taal-2Engels
TijdschriftJournal of Molecular Diagnostics
Volume16
Nummer van het tijdschrift4
Pagina's (van-tot)400-404
Aantal pagina's5
ISSN1525-1578
DOI's
StatusGepubliceerd - 2014

    Expertisedomeinen

  • Humans, Limit of Detection, RNA, Protozoan, RNA, Spliced Leader, Real-Time Polymerase Chain Reaction, Reverse Transcription, Trypanosoma brucei gambiense, Trypanosomiasis, African

ID: 1273034