1921
Volume 79, Issue 6
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645

Abstract

Traditional culture of parasites is labor-intensive and shows poor sensitivity. We evaluated microculture and novel miniculture methods for diagnosis of cutaneous leishmaniasis (CL). Consecutive patients who came to the Leishmaniasis Clinic, Hospital Nacional Cayetano Heredia, Lima, Peru, were enrolled. Lesion aspirates were cultured in traditional tubes containing Novy-MacNeal-Nicolle medium and in miniculture tubes (Eppendorf, Hamburg, Germany) and capillary tubes (microculture) containing RPMI 1640 medium containing 20% fetal bovine serum. The reference standard was positive results in two of four tests (smear, culture, polymerase chain reaction, or leishmanin skin test). Outcome measures were sensitivity and time to positivity. Fifty-five patients with 74 lesions were enrolled. Of 59 lesions that fulfilled reference criteria for CL, 50 were positive by microculture (sensitivity = 84.7%; = 0.001), 45 by miniculture (sensitivity = 76.3%; = 0.042), and 35 by traditional culture (sensitivity = 59.3%). Median time to positivity was three days by microculture and miniculture and five days by traditional culture ( < 0.001). Microculture and miniculture are sensitive and efficient means of diagnosing CL.

Loading

Article metrics loading...

/content/journals/10.4269/ajtmh.2008.79.847
2008-12-01
2017-09-26
Loading full text...

Full text loading...

/deliver/fulltext/14761645/79/6/0790847.html?itemId=/content/journals/10.4269/ajtmh.2008.79.847&mimeType=html&fmt=ahah

References

  1. Desjeux P, 2004. Leishmaniasis: current situation and new perspectives. Comp Immunol Microbiol Infect Dis 27 : 305–318.
  2. Pan American Health Organization, 2002. PAHO Basic Health Indicator Data Base. Peru. Available at: http://www.paho.org/English/DD/AIS/cp_604.htm. Accessed July 9, 2008.
  3. Arevalo J, Ramirez L, Adaui V, Zimic M, Tulliano G, Miranda-Verstegui C, Lazo M, Loayza-Muro R, De Doncker S, Mauer A, Chappuis F, Dujardin JC, Llanos-Cuentas A, 2007. Influence of Leishmania (Viannia) species on the response to antimonial treatment in patients with American tegumentary leishmaniasis. J Infect Dis 195 : 1846–1851.
  4. Lucas CM, Franke ED, Cachay MI, Tejada A, Cruz ME, Kreutzer RD, Barker DC, McCann SH, Watts DM, 1998. Geographic distribution and clinical description of leishmaniasis cases in Peru. Am J Trop Med Hyg 59 : 312–317.
  5. Santrich C, Segura I, Arias AL, Saravia NG, 1990. Mucosal disease caused by Leishmania braziliensis guyanensis. Am J Trop Med Hyg 42 : 51–55.
  6. Blum J, Desjeux P, Schwartz E, Beck B, Hatz C, 2004. Treatment of cutaneous leishmaniasis among travelers. J Antimicrob Chemother 53 : 158–166.
  7. Reithinger R, Dujardin JC, Louzir H, Pirmez C, Alexander B, Brooker S, 2007. Cutaneous leishmaniasis. Lancet Infect Dis 7 : 581–596.
  8. Murray HW, Berman JD, Davies CR, Saravia NG, 2005. Advances in leishmaniasis. Lancet 366 : 1561–1577.
  9. Singh S, 2006. New developments in diagnosis of leishmaniasis. Indian J Med Res 123 : 311–330.
  10. Tavares CA, Fernandes AP, Melo MN, 2003. Molecular diagnosis of leishmaniasis. Expert Rev Mol Diagn 3 : 657–667.
  11. Allahverdiyev AM, Uzun S, Bagirova M, Durdu M, Memisoglu HR, 2004. A sensitive new microculture method for diagnosis of cutaneous leishmaniasis. Am J Trop Med Hyg 70 : 294–297.
  12. Andresen K, Gaafar A, El-Hasan A, Ismail A, Dafalla M, Theander TG, Kharazmi A, 1996. Evaluation of the polymerase chain reaction in the diagnosis of cutaneous leishmaniasis due to Leishmania major. A comparison with direct microscopy of smears and sections from lesions. Trans R Soc Trop Med Hyg 90 : 133–135.
  13. Aviles H, Belli A, Armijos R, Monroy F, Harris E, 1999. PCR detection and identification of Leishmania parasites in clinical specimens in Ecuador: a comparison with direct microscopy of smears and sections from lesions. J Parasitol 85 : 181–187.
  14. Ramirez JR, Agudelo S, Muskus C, Alzate JF, Berberich C, Barker D, Velez ID, 2000. Diagnosis of cutaneous leishmaniasis in Colombia: the sampling site within lesions influences the sensitivity of parasitological diagnosis. J Clin Microbiol 38 : 3768–3773.
  15. Allahverdiyev AM, Bagirova M, Uzun S, Alabaz D, Aksaray N, Kocobas E, Koksal F, 2005. The value of a new microculture method for diagnosis of visceral leishmaniasis by using bone marrow and peripheral blood. Am J Trop Med Hyg 73 : 276–280.
  16. Ihalamulla RL, Rajapaksa US, Karunaweera ND, 2005. Micro-culture for the isolation of Leishmania parasites from cutaneous lesions: Sri Lankan experience. Ann Trop Med Parasitol 99 : 571–575.
  17. Ihalamulla RL, Rajapaksa US, Karunaweera ND, 2006. Micro-culture for the isolation of Leishmania, modified to increase efficacy: a follow-up to a previous study. Ann Trop Med Parasitol 100 : 87–89.
  18. Boggild AK, Miranda-Verastegui C, Espinosa D, Arevalo J, Adaui V, Tulliano G, Llanos-Cuentas A, Low DE, 2007. Evaluation of a microculture method for isolation of Leishmania parasites from cutaneous lesions of patients in Peru. J Clin Microbiol 45 : 3680–3684.
  19. Keegan F, Blum JJ, 1990. Effects of oxygen concentration on the intermediary metabolism of Leishmania major promastigotes. Mol Biochem Parasitol 39 : 235–245.
  20. Keegan F, Blum JJ, 1991. Changes in intracellular levels of fructose 2,6-bisphosphate and several glycolytic intermediates in Leishmania major promastigotes as a function of pO2. Mol Biochem Parasitol 47 : 161–166.
  21. Walsh MJ, Blum JJ, 1992. Effects of hypoxia and acute osmotic stress on intermediary metabolism in Leishmania promastigotes. Mol Biochem Parasitol 50 : 205–214.
  22. Chulay JD, Bryceson AD, 1983. Quantitation of amastigotes of Leishmania donovani in smears of splenic aspirates from patients with visceral leishmaniasis. Am J Trop Med Hyg 32 : 475–479.
  23. Sadeghian G, Momeni A, Siadat AH, Usefi P, 2006. Evaluation of leishmanin skin test and its relationship with the clinical form and duration of cutaneous leishmaniasis. Dermatol Online J 12 : 3.
  24. Sambrook J, Fritsch EF, Maniatis T, 1989. Molecular Cloning: A Laboratory Manual. Second edition. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press.
  25. Lopez M, Orrego C, Cangalaya M, Inga R, Arevalo J, 1993. Diagnosis of Leishmania via the polymerase chain reaction: a simplified procedure for field work. Am J Trop Med Hyg 49 : 348–356.
  26. Romero GA, Vinitius de Farias Guerra M, Gomes Paes M, de Oliviera Macedo V, 2001. Comparison of cutaneous leishmaniasis due to Leishmania (Viannia) braziliensis and L. (V.) guyanensis in Brazil: clinical findings and diagnostic approach. Clin Infect Dis 32 : 1304–1312.
  27. Tojal da Silva AC, Cupolillo E, Volpini AC, Almeida R, Sierra Romero GA, 2006. Species diversity causing human cutaneous leishmaniasis in Rio Branco, state of Acre, Brazil. Trop Med Int Health 11 : 1388–1398.
  28. Blum JT, Junghanss T, Hatz C, 1994. Erroneous tracks in the diagnosis of cutaneous and mucocutaneous leishmaniasis. Schweiz Rundsch Med Prax 83 : 1025–1029.
  29. Jones TC, Johnson WD, Barretto AC, Lago E, Badar R, Cerf B, Reed SG, Netto EM, Tada MS, Franca TF, 1987. Epidemiology of American cutaneous leishmaniasis due to Leishmania braziliensis braziliensis. J Infect Dis 156 : 73–83.
  30. Marsden PD, 1986. Mucosal leishmaniasis (espundia‘ Escomel, 1911). Trans R Soc Trop Med Hyg 80 : 859–876.
  31. Llanos-Cuentas A, Tulliano G, Araujo-Castillo R, Miranda-Verstagui C, Santamaria-Castrellon G, Ramirez L, Lazo M, De Doncker S, Boelaert M, Robays J, Dujardin JC, Arevalo J, Chappuis F, 2008. Clinical and parasite species risk factors for pentavalent antimonial treatment failure in cutaneous leishmaniasis in Peru. Clin Infect Dis 46 : 223–231.
  32. Bensoussan E, Nasereddin A, Jonas F, Schnur LF, Jaffe CL, 2006. Comparison of PCR assays for diagnosis of cutaneous leishmaniasis. J Clin Microbiol 44 : 1435–1439.
  33. Faber WR, Oskam L, van Gool T, Kroon NC, Knegt-Junk KJ, Hofwegen H, van der Wal AC, Kager PA, 2003. Value of diagnostic techniques for cutaneous leishmaniasis. J Am Acad Dermatol 49 : 70–74.
  34. Manzur A, Bari A, 2006. Sensitivity of leishmanin skin test in patients of acute cutaneous leishmaniasis. Dermatol Online J 12 : 2.
  35. Weina PJ, Neafie RC, Wortmann G, Polhemus M, Aronson NE, 2004. Old World leishmaniasis; an emerging infection among deployed US military and civilian workers. Clin Infect Dis 39 : 1674–1680.
  36. Banoo S, Bell D, Bossuyt P, Herring A, Mabey D, Poole F, Smith PG, Sriram N, Wongsrichanalai C, Linke R, O’Brien R, Perkins M, Cunningham J, Matsoso P, Nathanson CM, Olliaro P, Peeling RW, Ramsay A, 2007. Evaluating diagnostic tests for infectious diseases: general principles. Nat Rev Microbiol 5 (Suppl): S17–S27.
  37. Croft SL, Sundar S, Fairlamb AH, 2006. Drug resistance in leishmaniasis. Clin Microbiol Rev 19 : 111–126.
http://instance.metastore.ingenta.com/content/journals/10.4269/ajtmh.2008.79.847
Loading
/content/journals/10.4269/ajtmh.2008.79.847
Loading

Data & Media loading...

  • Received : 14 Jul 2008
  • Accepted : 27 Aug 2008

Most Cited This Month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error