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Development of a Reverse Transcriptase Loop-Mediated Isothermal Amplification (LAMP) Assay for the Sensitive Detection of Leishmania Parasites in Clinical Samples

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  • 1 Koninklijk Instituut voor de Tropen (KIT) Biomedical Research, Amsterdam, The Netherlands; Faculty of Medicine, University of Khartoum, Khartoum, Sudan

Here we describe a generic, reverse transcriptase-loop-mediated isothermal amplification (RT-LAMP) assay, for the identification of Leishmania species from clinical samples. LAMP is an isothermal reaction recently developed as a point-of-care diagnostic tool. Primers were designed in the conserved region of the 18S ribosomal RNA (rRNA) gene; amplification was visualized by the pre-amplification addition of fluorescent detection reagent (FDR) and a simple UV lamp. By using a reverse-transcriptase step, the system detected infections between 10 and 100 parasites per mL. The assay was tested on a range of nucleic acid extracts from Leishmania species, visceral leishmaniasis (VL) patients from Sudan, and cutaneous leishmaniasis (CL) patients from Suriname. The sensitivity of RT-LAMP from the blood of VL patients was 83% (N = 30) compared with microscopy of bone-marrow and lymph-node aspirates; for CL patients the observed sensitivity was 98% (N = 43). The potential to use LAMP as a diagnostic tool for leishmaniasis is discussed.

Author Notes

*Address correspondence to Emily R. Adams, Koninklijk Instituut voor de Tropen (KIT), KIT Biomedical Research, Meibergdreef 39, 1105 AZ Amsterdam, The Netherlands. E-mail: e.adams@kit.nl

Financial support: KIT. Clinical material from Sudan was collected within the framework of a collaborative research project sponsored by the European community FP6 call - priority INCO-DEV - TRYLEIDIAG, contract 015379. Collection of samples from Suriname was funded by the Netherlands Organization for Scientific Research – WOTRO Science for Global Development contract W96-210.

Authors' addresses: Emily R. Adams, Gerard J. Schoone, and Henk D. F. H. Schallig, Koninklijk Instituut voor de Tropen (KIT), KIT Biomedical Research, Amsterdam, The Netherlands, E-mails: e.adams@kit.nl, g.schoone@kit.nl, and h.schallig@kit.nl. Al Farazdag Ageed and Sayda El Safi, Faculty of Medicine, University of Khartoum, Khartoum, Sudan, E-mails: elfarazdega@yahoo.com and shelsafi@yahoo.com.

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