Diagnosis of Visceral Leishmaniasis Using Peripheral Blood Microscopy in Ethiopia: A Prospective Phase-III Study of the Diagnostic Performance of Different Concentration Techniques Compared to Tissue Aspiration

Ermias Diro University of Gondar, Gondar, Ethiopia.
Institute of Tropical Medicine, Antwerp, Belgium.

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Cedric P. Yansouni J.D. MacLean Centre for Tropical Diseases at McGill University, Montreal, Canada.

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Yegnasew Takele University of Gondar, Gondar, Ethiopia.

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Bewketu Mengesha University of Gondar, Gondar, Ethiopia.

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Lutgarde Lynen Institute of Tropical Medicine, Antwerp, Belgium.

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Asrat Hailu School of Medicine, Addis Ababa University College of Health Sciences, Addis Ababa, Ethiopia.

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Johan van Griensven Institute of Tropical Medicine, Antwerp, Belgium.

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Marleen Boelaert Institute of Tropical Medicine, Antwerp, Belgium.

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Philippe Büscher Institute of Tropical Medicine, Antwerp, Belgium.

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DOI:
https://doi.org/10.4269/ajtmh.16-0362
Volume/Issue:
Volume 96: Issue 1
Page(s):
190–196
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Visceral leishmaniasis (VL) is a fatal parasitic disease. Unfortunately, diagnosis of VL in east Africa currently relies on aspiration of tissue from the spleen or bone marrow, which is painful and potentially dangerous. We sought to determine whether peripheral blood could be used instead of invasive tissue aspirates to diagnose VL, using three parasite concentration techniques. Three hundred and one consecutive people suspected of having VL were recruited. Compared with microscopy of tissue aspirates, the diagnostic accuracy of peripheral blood microscopy was as follows: whole blood thin smear sensitivity 1.5% (95% confidence interval [CI] 0.0–8.3) and specificity 100% (95% CI 76.8–100); buffy-coat smear sensitivity 19.5% (95% CI 14.3–25.6) and specificity 98.9% (95% CI 94.1–100); peripheral blood mononuclear cell (PBMC) smear sensitivity 33.7% (95% CI 27.3–40.5) and specificity 95.7% (95% CI 89.6–98.6). Sensitivity of PBMC smears was significantly higher in human immunodeficiency virus (HIV)-coinfected patients (N = 48/301); two-sample test of proportions, P = 0.0097; sensitivity 55.9% (95% CI 37.9–72.8) and specificity 92.9% (95% CI 66.1–99.8), and correlated with the degree of parasite load in the tissue. Combining the results from smears of both PBMC and buffy coat yielded a sensitivity and specificity of 67.6% (95% CI 49.1–82.6) and 92.9% (95% CI 66.1–99.8), respectively, in HIV-coinfected patients. In this setting, VL could be ruled-in with peripheral blood microscopy in a substantial number of VL suspects and may reduce the number of tissue aspirations performed, particularly in HIV-coinfected patients. More sensitive and logistically feasible methods than light microscopy are needed to detect Leishmania donovani parasites present in blood.

Author Notes

* Address correspondence to Cédric Yansouni, Division of Infectious Diseases, Department of Microbiology, J.D. MacLean Centre for Tropical Diseases at McGill University, McGill University Health Centre, 1001 Décarie Bouelvard, Room EM3.3242, Montreal, QC, H4A 3J1, Canada. E-mail: cedric.yansouni@mcgill.ca
† These authors contributed equally to this work.

Financial support: This study was supported by a grant from AIDS Mecenaat at Institute of Tropical Medicine (ITM). Ermias Diro has received a PhD scholarship granted from the Belgian Directorate General for Development Cooperation under the ITM-DGDC framework agreement FA-III and from the European Union Seventh Framework Programme (FP7/2007-2013) under grant agreement no. 305178 via AfriCoLeish project. Cedric Yansouni holds a “Chercheur-boursier clinicien” career award from the Fonds de recherche du Québec—Santé (FRQS).

Authors' addresses: Ermias Diro, University of Gondar, Gondar, Ethiopia, and Institute of Tropical Medicine, Antwerp, Belgium, E-mail: ermi_diro@yahoo.com. Cedric Yansouni, J.D. MacLean Centre for Tropical Diseases at McGill University, Montreal, Canada, E-mail: cedric.yansouni@mcgill.ca. Yegnasew Takele and Bewketu Mengesha, University of Gondar, Gondar, Ethiopia, E-mails: yegnasew77@yahoo.com and bewketmd@yahoo.com. Lutgarde Lynen, Johan van Griensven, Marleen Boelaert, and Philippe Büscher, Institute of Tropical Medicine, Antwerp, Belgium, E-mails: llynen@itg.be, jvangriensven@itg.be, mboelaert@itg.be, and pbuscher@itg.be. Asrat Hailu, School of Medicine, Addis Ababa University College of Health Sciences, Addis Ababa, Ethiopia, E-mail: hailu_a2004@yahoo.com.

Copyright:
© The American Society of Tropical Medicine and Hygiene 2017
Received:
06 May 2016
|
Accepted:
25 Sep 2016
|
Published Online:
11 Jan 2017
Cover The American Journal of Tropical Medicine and Hygiene
The American Journal of Tropical Medicine and Hygiene
Print ISSN:
0002-9637
Online ISSN:
1476-1645
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