1887

Abstract

Introduction:

Retrospective molecular identification of parasites in two patients with visceral leishmaniasis (VL) previously treated in Serbia was carried out. DNA was isolated from unstained bone marrow smears (BMSs) kept for 11 and 8 years. Genus-specific real-time PCR was combined with conventional PCR and sequencing for detection and species identification.

Case presentation:

In 2003, a 40-year-old Serbian male was admitted to the Clinical Centre of Serbia (CCS) with fever, sweating, fatigue and splenomegaly, which developed over a period of 7 weeks. He had frequently travelled around Europe. VL was confirmed by microscopy of Giemsa-stained BMS. Treatment by pentavalent antimonials was successfully completed. Two years later, the patient developed post-kala-azar dermal leishmaniasis. Treatment resulted in symptom resolution. Later on, was identified as the causative agent of the VL by sequencing of the ITS (internal transcribed spacer) region; mixed spp. infection could not be excluded. In 2006, a 33-year-old female from Vojvodina, Serbia, with pre-existing diabetes mellitus and chronic meningoencephalitis and a history of frequent visits to the Montenegrin seacoast, was admitted to the CCS with fever, pancytopenia and moderate hepatosplenomegaly. A stained BMS revealed abundant amastigotes. Indirect haemagglutination analysis was positive with a titre of 1 : 2048, and a rapid dipstick rK39 test was also positive. Treatment by liposomal amphotericin B was successful; however, shortly after, the patient developed neural infection and pneumonia and died. The causative agent was identified as .

Conclusion:

Molecular diagnosis of VL and species delineation using DNA from unstained BMSs stored for several years is possible.

Loading

Article metrics loading...

/content/journal/jmmcr/10.1099/jmmcr.0.005063
2016-09-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/jmmcr/3/5/jmmcr005063.html?itemId=/content/journal/jmmcr/10.1099/jmmcr.0.005063&mimeType=html&fmt=ahah

References

  1. Beatović V. 2010; The Institute of Public Health of Montenegro, The Ministry ofHealth of Montenegro, WHO Exploratory Meeting on Leishmaniasis in the Balkan Countries, Dubrovnik, Croatia, 10–12 February 2010. Available on: http://www.who.int/leishmaniasis/resources/MONTENEGRO.pdf
  2. Brustoloni Y. M., Lima R. B., Da Cunha R. V., Dorval M. E., Oshiro E. T., De Oliveira A. L., Pirmez C. 2007; Sensitivity and specificity of polymerase chain reaction in Giemsa-stained slides for diagnosis of visceral leishmaniasis in children. Mem Inst Oswaldo Cruz 102:497–500 [View Article][PubMed]
    [Google Scholar]
  3. Cirović D., Chochlakis D., Tomanović S., Sukara R., Penezić A., Tselentis Y., Psaroulaki A. 2014; Presence of Leishmania and Brucella species in the golden jackal Canis aureus in Serbia. Biomed Res Int 2014:728516 [View Article][PubMed]
    [Google Scholar]
  4. Cortes S., Rolão N., Ramada J., Campino L. 2004; PCR as a rapid and sensitive tool in the diagnosis of human and canine leishmaniasis using Leishmania donovani s.l.-specific kinetoplastid primers. Trans R Soc Trop Med Hyg 98:12–17 [View Article][PubMed]
    [Google Scholar]
  5. Da Silva E. S., Gontijo C. M., Pacheco R. S., Brazil R. P. 2004; Diagnosis of human visceral leishmaniasis by PCR using blood samples spotted on filter paper. Genet Mol Res 3:251–257[PubMed]
    [Google Scholar]
  6. Dakic Z. D., Pelemis M. R., Stevanovic G. D., Poluga J. L., Lavadinovic L. S., Milosevic I. S., Indjic N. K., Ofori-Belic I. V., Pavlovic M. D. 2009; Epidemiology and diagnostics of visceral leishmaniasis in Serbia. Clin Microbiol Infect 15:1173–1176 [View Article][PubMed]
    [Google Scholar]
  7. Das N. K., Singh S. K., Ghosh S., Sarkar A., Mukhopadhyay D., Roy S., Ganguly D. N., Barbhuiya J. N., Saha B., Chatterjee M. 2011; Case series of misdiagnosis with rK39 strip test in Indian leishmaniasis. Am J Trop Med Hyg 84:688–691 [View Article][PubMed]
    [Google Scholar]
  8. De Ruiter C. M., Van der Veer C., Leeflang M. M., Deborggraeve S., Lucas C., Adams E. R. 2014; Molecular tools for diagnosis of visceral leishmaniasis: systematic review and meta-analysis of diagnostic test accuracy. J Clin Microbiol 52:3147–3155 [View Article][PubMed]
    [Google Scholar]
  9. Durda A. 2010; WHO Exploratory Meeting on Leishmaniasis in the Balkan Countries. Dubrovnik, Croatia, 10–12 February 2010 http://www.who.int/leishmaniasis/resources/BOSNIA.pdf
  10. Ganguly S., Das N. K., Barbhuiya J. N., Chatterjee M. 2010; Post-kala-azar dermal leishmaniasis - an overview. Int J Dermatol 49:921–931 [View Article][PubMed]
    [Google Scholar]
  11. Gebhardt M., Ertas B., Falk T. M., Blödorn-Schlicht N., Metze D., Böer-Auer A. 2015; Fast, sensitive and specific diagnosis of infections with Leishmania spp. in formalin-fixed, paraffin-embedded skin biopsies by cytochrome b polymerase chain reaction. Br J Dermatol 173:1239–1249 [View Article][PubMed]
    [Google Scholar]
  12. Gradoni L. 2013; Epidemiological surveillance of leishmaniasis in the European Union: operational and research challenges. Euro Surveill 18:20539 [View Article][PubMed]
    [Google Scholar]
  13. Haldar J. P., Saha K. C., Ghose A. C. 1981; Serological profiles in Indian post kala-azar dermal leishmaniasis. Trans R Soc Trop Med Hyg 75:514–517 [View Article][PubMed]
    [Google Scholar]
  14. Institute of Public Health of Serbia 2009; Report on Infectious Diseases in the Republic of Serbia in 2008, p. 28 (in Serbian). Available on: http://www.batut.org.rs/index.php?content=299
  15. Institute of Public Health of Serbia 2015; Report on Infectious Diseases in the Republic of Serbia in 2014, p. 63 (in Serbian). Available on: http://www.batut.org.rs/download/izvestaji/Izvestaj%20o%20zaraznim%20bolestima%202014.pdf
  16. Ivović V., Depaquit J., Léger N., Urano A., Papadopoulos B. 2004; Sandflies (Diptera: Psychodidae) in the bar area of Montenegro (Yugoslavia). 2. Presence of promastigotes in Phlebotomus neglectus and first record of P. kandelakii . Ann Trop Med Parasitol 98:425–427 [View Article][PubMed]
    [Google Scholar]
  17. Lanús E. C., Piñero J. E., González A. C., Valladares B., De Grosso M. L., Salomón O. D. 2005; Detection of Leishmania braziliensis in human paraffin-embedded tissues from Tucumán, Argentina by polymerase chain reaction. Mem Inst Oswaldo Cruz 100:187–192 [View Article][PubMed]
    [Google Scholar]
  18. Lipoldová M., Demant P. 2006; Genetic susceptibility to infectious disease: lessons from mouse models of leishmaniasis. Nat Rev Genet 7:294–305 [View Article][PubMed]
    [Google Scholar]
  19. Mahmoudzadeh-Niknam H., Abrishami F., Doroudian M., Moradi M., Alimohammadian M., Parvizi P., Hatam G., Mohebali M., Khalaj V. 2011; The problem of mixing up of Leishmania isolates in the laboratory: suggestion of ITS1 gene sequencing for verification of species. Iran J Parasitol 6:41–48[PubMed]
    [Google Scholar]
  20. Marjanovic G., Miladinovic-Tasic N., Gabrielli S., Otasevic S., Popovic-Dragonjic L., Kocic B., Arsic-Arsenijevic V., Tadic L., Cancrini G. 2012; First case of visceral leishmaniosis/HIV coinfection in Nis - southeastern Serbia. Archives Biological Sci 64:1271–1276 [View Article]
    [Google Scholar]
  21. Monge-Maillo B., Norman F. F., Cruz I., Alvar J., López-Vélez R. 2014; Visceral leishmaniasis and HIV coinfection in the Mediterranean region. PLoS Negl Trop Dis 21:e3021 [View Article]
    [Google Scholar]
  22. Peacock C. S., Seeger K., Harris D., Murphy L., Ruiz J. C., Quail M. A., Peters N., Adlem E., Tivey A. et al. 2007; Comparative genomic analysis of three Leishmania species that cause diverse human disease. Nat Genet 39:839–847 [View Article][PubMed]
    [Google Scholar]
  23. Savić S., Vidić B., Grgić Z., Potkonjak A., Spasojevic L. 2014; Emerging vector-borne diseases - incidence through vectors. Front Public Health 2:267 [View Article][PubMed]
    [Google Scholar]
  24. Singh S., Sharma U., Mishra J. 2011; Post-kala-azar dermal leishmaniasis: recent developments. Int J Dermatol 50:1099–1108 [View Article][PubMed]
    [Google Scholar]
  25. Šiško-Kraljević K., Jerončić A., Mohar B., Punda-Polić V. 2013; Asymptomatic Leishmania infantum infections in humans living in endemic and non-endemic areas of Croatia, 2007 to 2009. Euro Surveill 18:20533 [View Article][PubMed]
    [Google Scholar]
  26. Vaselek S., Savić S., Di Muccio T., Erisoz Kasap O., Gradoni L., Alten B., Petrić D. 2015; Possible re-emergence of leishmaniasis in Serbia. In Abstracts of the 2nd Conference on Neglected Vectors and Vector-Borne Diseases with MC and WG Meetings of the COST Action TD1303, Izmir, Turkey, 31 March–2 April 2015, p. 4 http://eurnegvec.org/2ac_abstractbook.pdf
  27. Zijlstra E. E. 2014; PKDL and other dermal lesions in HIV coinfected patients with leishmaniasis: review of clinical presentation in relation to immune responses. PLoS Negl Trop Dis 20:e3258
    [Google Scholar]
  28. Zijlstra E. E., Musa A. M., Khalil E. A., El-Hassan I. M., El-Hassan A. M. 2003; Post-kala-azar dermal leishmaniasis. Lancet Infect Dis 3:87–98 [View Article][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmmcr/10.1099/jmmcr.0.005063
Loading
/content/journal/jmmcr/10.1099/jmmcr.0.005063
Loading

Data & Media loading...

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