title: Usefulness of PCR method for detection of Leishmania in Poland.
abstract: Leishmania parasites are the etiological agents of leishmaniosis, with severe course and often fatal prognosis, and the global number of cases has increased in recent decades. The gold standards for the diagnosis of leishmaniosis are microscopic examinations and culture in vitro of the different clinical specimens. The sensitivity of these methods is insufficient. Recent development in specific and sensitive molecular methods (PCR) allows for detection as well as identification of the parasite species (subspecies). The aim of the study was to estimate the usefulness of molecular methods (PCR) for detection of Leishmania species and consequently for the implementation of such methods in routine diagnostics of leishmaniosis in Polish patients returning from endemic areas of the disease. In our investigations we used 54 known Leishmania positive DNA templates (from culture and clinical specimens) received from the CDC (Atlanta, GA, USA). Moreover, 25 samples of bone marrow, blood or other tissues obtained from 18 Polish individuals suspected of leishmaniosis were also examined. In PCR we used two pairs of primers specific to the conserved region of Leishmania kinetoplast DNA (kDNA) minicircle (13A/13B and F/R). Using these primers we obtained amplicons in all DNA templates from the CDC and in three Polish patients suspected for Leishmania infection. In one sample from among these cases we also obtained positive results with DNA isolated from a blood specimen which was previously negative in microscopic examinations.
authors.affiliation: Department of Tropical Medicine and Parasitology, Interfaculty Institute of Maritime and Tropical Medicine in Gdynia, Medical University of Gdańsk, Gdynia, Poland. pemyjak@gumed.edu.pl
authors.forename: Przemysław
authors.initials: P
authors.lastname: Myjak
authors.forename: Joanna
authors.initials: J
authors.lastname: Szulta
authors.forename: Marcos E
authors.initials: ME
authors.lastname: de Almeida
authors.forename: Alexandre J
authors.initials: AJ
authors.lastname: da Silva
authors.forename: Francis
authors.initials: F
authors.lastname: Steurer
authors.forename: Anna
authors.initials: A
authors.lastname: Lass
authors.forename: Halina
authors.initials: H
authors.lastname: Pietkiewicz
authors.forename: Wacław L
authors.initials: WL
authors.lastname: Nahorski
authors.forename: Jolanta
authors.initials: J
authors.lastname: Goljan
authors.forename: Józef
authors.initials: J
authors.lastname: Knap
authors.forename: Beata
authors.initials: B
authors.lastname: Szostakowska
mesh_terms: D016054:DNA, Protozoan; D006801:Humans; D007891:Leishmania / Q000302:isolation & purification*; D007896:Leishmaniasis / Q000175:diagnosis* / Q000453:epidemiology*; D011044:Poland / Q000453:epidemiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D057236:Travel Medicine
chemical_list: D016054:DNA, Protozoan