Volume 96, Issue 1
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645



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 ( = 48/301); two-sample test of proportions, = 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 parasites present in blood.


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  1. Alvar J, Velez ID, Bern C, Herrero M, Desjeux P, Cano J, Jannin J, den Boer M, , 2012. Leishmaniasis worldwide and global estimates of its incidence. PLoS One 7: e35671.[Crossref] [Google Scholar]
  2. van Griensven J, Diro E, , 2012. Visceral leishmaniasis. Infect Dis Clin North Am 26: 309322.[Crossref] [Google Scholar]
  3. Alvar J, Bashaye S, Argaw D, Cruz I, Aparicio P, Kassa A, Orfanos G, Parreno F, Babaniyi O, Gudeta N, Canavate C, Bern C, , 2007. Kala-azar outbreak in Libo Kemkem, Ethiopia: epidemiologic and parasitologic assessment. Am J Trop Med Hyg 77: 275282. [Google Scholar]
  4. Bern C, Maguire JH, Alvar J, , 2008. Complexities of assessing the disease burden attributable to leishmaniasis. PLoS Negl Trop Dis 2: e313.[Crossref] [Google Scholar]
  5. Boelaert M, Verdonck K, Menten J, Sunyoto T, van Griensven J, Chappuis F, Rijal S, , 2014. Rapid tests for the diagnosis of visceral leishmaniasis in patients with suspected disease. Cochrane Database Syst Rev 6: CD009135. [Google Scholar]
  6. Cunningham J, Hasker E, Das P, El Safi S, Goto H, Mondal D, Mbuchi M, Mukhtar M, Rabello A, Rijal S, Sundar S, Wasunna M, Adams E, Menten J, Peeling R, Boelaert M, , 2012. A global comparative evaluation of commercial immunochromatographic rapid diagnostic tests for visceral leishmaniasis. Clin Infect Dis 55: 13121319.[Crossref] [Google Scholar]
  7. Medrano FJ, Canavate C, Leal M, Rey C, Lissen E, Alvar J, , 1998. The role of serology in the diagnosis and prognosis of visceral leishmaniasis in patients coinfected with human immunodeficiency virus type-1. Am J Trop Med Hyg 59: 155162. [Google Scholar]
  8. ter Horst R, Collin SM, Ritmeijer K, Bogale A, Davidson RN, , 2008. Concordant HIV infection and visceral leishmaniasis in Ethiopia: the influence of antiretroviral treatment and other factors on outcome. Clin Infect Dis 46: 17021709.[Crossref] [Google Scholar]
  9. Diro E, Lynen L, Ritmeijer K, Boelaert M, Hailu A, van Griensven J, , 2014. Visceral Leishmaniasis and HIV coinfection in east Africa. PLoS Negl Trop Dis 8: e2869.[Crossref] [Google Scholar]
  10. Gidwani K, Picado A, Ostyn B, Singh SP, Kumar R, Khanal B, Lejon V, Chappuis F, Boelaert M, Sundar S, , 2011. Persistence of Leishmania donovani antibodies in past visceral leishmaniasis cases in India. Clin Vaccine Immunol 18: 346348.[Crossref] [Google Scholar]
  11. Boelaert M, el Safi S, Goetghebeur E, Gomes-Pereira S, Le Ray D, Van der Stuyft P, , 1999. Latent class analysis permits unbiased estimates of the validity of DAT for the diagnosis of visceral leishmaniasis. Trop Med Int Health 4: 395401.[Crossref] [Google Scholar]
  12. Chemli J, Abroug M, Fathallah A, Abroug S, Ben SM, Harbi A, , 2006. Contribution of leukoconcentration in the diagnosis of Kala-azar in Tunisia [in French]. Med Mal Infect 36: 390392.[Crossref] [Google Scholar]
  13. Deniau M, Canavate C, Faraut-Gambarelli F, Marty P, , 2003. The biological diagnosis of leishmaniasis in HIV-infected patients. Ann Trop Med Parasitol 97 (Suppl 1): 115133.[Crossref] [Google Scholar]
  14. Izri MA, Robineau M, Petithory JC, Rousset JJ, , 1993. Visceral leishmaniasis. Parasitological diagnosis by leukocyte concentration [in French]. Presse Med 22: 1010. [Google Scholar]
  15. Izri MA, Deniau M, Briere C, Rivollet D, Petithory JC, Houin R, Rousset JJ, , 1996. Leishmaniasis in AIDS patients: results of leukocytoconcentration, a fast biological method of diagnosis. Bull World Health Organ 74: 9193. [Google Scholar]
  16. Boelaert M, Bhattacharya S, Chappuis F, el Safi S, Hailu A, Mondal D, Rijal S, Sundar S, Wasunna M, Peeling RW, , 2007. Evaluation of rapid diagnostic tests: visceral leishmaniasis. Nat Rev Microbiol 5: S30S39.[Crossref] [Google Scholar]
  17. World Health Organization, 2010. Control of the Leishmaniases. Report of a meeting of the WHO Expert Committee on the Control of Leishmaniases, Geneva, March 22–26, 2010. Geneva, Switzerland: WHO Technical Report Series 949, World Health Organization. [Google Scholar]
  18. Petithory JC, Ardoin F, Ash LR, Vandemeulebroucke E, Galeazzi G, Dufour M, Paugam A, , 1997. Microscopic diagnosis of blood parasites following a cytoconcentration technique. Am J Trop Med Hyg 57: 637642. [Google Scholar]
  19. Boelaert M, Rijal S, Regmi S, Singh R, Karki B, Jacquet D, Chappuis F, Campino L, Desjeux P, Le Ray D, Koirala S, Van der Stuyft P, , 2004. A comparative study of the effectiveness of diagnostic tests for visceral leishmaniasis. Am J Trop Med Hyg 70: 7277. [Google Scholar]
  20. Diro E, Lynen L, Mohammed R, Boelaert M, Hailu A, van Griensven J, , 2014. High parasitological failure rate of visceral leishmaniasis to sodium stibogluconate among HIV co-infected adults in Ethiopia. PLoS Negl Trop Dis 8: e2875.[Crossref] [Google Scholar]
  21. Green TA, Black CA, Johnson RE, , 2001. In defense of discrepant analysis. J Clin Epidemiol 54: 210215.[Crossref] [Google Scholar]
  22. Lipman HB, Astles JR, , 1998. Quantifying the bias associated with use of discrepant analysis. Clin Chem 44: 108115. [Google Scholar]
  23. Schachter J, , 2001. In defense of discrepant analysis. J Clin Epidemiol 54: 211212.[Crossref] [Google Scholar]

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  • Received : 06 May 2016
  • Accepted : 25 Sep 2016
  • Published online : 11 Jan 2017

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