Microscopy Research, 2015, 3, 17-25 Published Online April 2015 in SciRes. http://www.scirp.org/journal/mr http://dx.doi.org/10.4236/mr.2015.32003 How to cite this paper: Mitashi, P., et al. (2015) Improved Detection of Sleeping Sickness Cases by LED Fluorescence Mi- croscopy: Evidence from a Prospective Multi-Centric Study in the Democratic Republic of the Congo. Microscopy Research, 3, 17-25. http://dx.doi.org/10.4236/mr.2015.32003 Improved Detection of Sleeping Sickness Cases by LED Fluorescence Microscopy: Evidence from a Prospective Multi-Centric Study in the Democratic Republic of the Congo Patrick Mitashi 1 , Pascal Lutumba 1 , Crispin Lumbala 2 , Paul Bessell 3 , Sylvain Biéler 3* , Joseph Mathu Ndung’u 3 1 Tropical Medicine Department, Faculty of Medicine, University of Kinshasa, Kinshasa, Democratic Republic of the Congo 2 Programme National de Lutte contre la Trypanosomiase Humaine Africaine (PNLTHA), Croisement de l’Avenue de la Libération et du Boulevard Triomphal 1, Kinshasa, Democratic Republic of the Congo 3 Foundation for Innovative New Diagnostics (FIND), Campus Biotech, Chemin des Mines 9, Geneva, Switzerland Email: drmitashi@yahoo.fr , pascal_lutumba@yahoo.fr , crispinlumbala@gmail.com , paul.bessell@finddiagnostics.org , * sylvain.bieler@finddiagnostics.org , joseph.ndungu@finddiagnostics.org Received 20 January 2015; accepted 27 March 2015; published 3 April 2015 Copyright © 2015 by authors and Scientific Research Publishing Inc. This work is licensed under the Creative Commons Attribution International License (CC BY). http://creativecommons.org/licenses/by/4.0/ Abstract Background: Confirmatory diagnosis of Trypanosoma brucei gambiense human African trypano- somiasis (HAT) is based on demonstration of parasites by microscopy. However, the sensitivity of routine microscopy methods is very low, and many cases are missed and left untreated. A clinical study was conducted in the Democratic Republic of the Congo to evaluate the accuracy of improved microscopy methods in diagnosis of HAT. These included examination by fluorescence microscopy (FM) of acridine orange (AO) stained smears of whole blood and smears made following a new procedure for concentrating trypanosomes by selective lysis of red blood cells (RBC). Methodol- ogy/Principal Findings: Venous blood was collected from 213 HAT cases, 101 HAT suspects and 95 controls and used to determine the accuracy of four microscopy methods: bright field microscopy of Giemsa-stained thick blood smears, FM of AO-stained thick blood smears, FM of AO-stained thick blood smears prepared after RBC lysis and concentration, and FM of AO-stained thin blood smears prepared after RBC lysis and concentration. The sensitivity of FM using thick blood smears stained with AO was 3 times higher than bright field microscopy using Giemsa-stained thick blood smears * Corresponding author.