{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,4]],"date-time":"2025-11-04T23:20:58Z","timestamp":1762298458157},"reference-count":28,"publisher":"Microbiology Society","issue":"5","license":[{"start":{"date-parts":[[2016,9,1]],"date-time":"2016-09-01T00:00:00Z","timestamp":1472688000000},"content-version":"unspecified","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2016,9,1]]},"abstract":"<jats:sec sec-type=\"intro\">\n                  <jats:title>Introduction:<\/jats:title>\n                  <jats:p>Retrospective molecular identification of <jats:italic>Leishmania <\/jats:italic>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.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Case presentation:<\/jats:title>\n                  <jats:p>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, <jats:italic>Leishmania infantum<\/jats:italic> was identified as the causative agent of the VL by sequencing of the ITS (internal transcribed spacer) region; mixed <jats:italic>Leishmania<\/jats:italic> 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 <jats:italic>Leishmania<\/jats:italic> amastigotes. Indirect haemagglutination analysis was positive with a titre of 1\u2009:\u20092048, 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 <jats:italic>L. infantum<\/jats:italic>.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec sec-type=\"conclusions\">\n                  <jats:title>Conclusion:<\/jats:title>\n                  <jats:p>Molecular diagnosis of VL and species delineation using DNA from unstained BMSs stored for several years is possible.<\/jats:p>\n               <\/jats:sec>","DOI":"10.1099\/jmmcr.0.005063","type":"journal-article","created":{"date-parts":[[2016,8,18]],"date-time":"2016-08-18T11:12:23Z","timestamp":1471518743000},"source":"Crossref","is-referenced-by-count":2,"title":["Retrospective PCR-based species identification of Leishmania in two patients with visceral leishmaniasis in Serbia"],"prefix":"10.1099","volume":"3","author":[{"given":"Zorica","family":"Daki\u0107","sequence":"first","affiliation":[{"name":"Parasitological Laboratory, Department of Microbiology, Clinical Centre of Serbia, Bulevar Oslobodjenja 16, 11000 Belgrade, 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