Akademik Veri Yönetim Sistemi
Tezin Türü: Doktora
Tezin Yürütüldüğü Kurum: İstanbul Üniversitesi-Cerrahpaşa, Cerrahpaşa Tıp Fakültesi, Temel Tıp Bilimleri Bölümü, Türkiye
Tezin Onay Tarihi: 2017
Tezin Dili: Türkçe
Öğrenci: SERHAT SİREKBASAN
Danışman: Erdal Polat
Özet:
Sirekbasan S. Diagnosis of Leishmania and Determination of Types on Clinical
Samples by Using High Resolution Melting Analysis (HRMA) Method. İstanbul
University, Institute of Health Science, Department of Medical Microbiology. Doctoral
Thesis. İstanbul. 2017.
Leishmaniosis is a group of clinical diseases caused by the Leishmania parasite, which
is transmitted by the bite of infected Phlebotomus and Lutzomyia sand flies, and is more
common in rural areas. Leishmaniosis, which is endemic in 88 countries (22 in the new
world and 66 in the old world) on 4 continents, is ranked second on the list of six
important tropical diseases.
It is very important that parasite species are identified quickly and correctly for proper
treatment planning and disease prevention. Because the course and severity of the
disease are not the same and the treatment regimes vary in the different species that
constitute the same clinical picture. According to information found in the book, the
species causing leishmaniosis in our country are mostly L. tropica and L. infantum.
However, about 3 million refugees have escaped to our country from the civil war in
Syria since December 2011, and thus the new species and strains resistant to
pentavalent antimony derivatives have been added to these species. Therefore, it is
important to identify the causative agent of leishmaniosis and, if necessary, the drug
resistance, in terms of treatment of the disease and taking the necessary precautions.
We can say that only Leishmania is seen and produced via direct microscopy and
culture methods routinely used in the diagnosis of leishmaniosis. Molecular methods are
needed to determine the type of Leishmania and whether it is resistant to drugs.
Therefore, the HRMA method, which is molecularly simple, fast and cost effective, was
used in our study. A total of 85 patients, 6 with suspected VL and 79 with suspected
CL, were examined in the study. The slides prepared from samples were stained with
Giemsa's stain and were examined under the light microscope. Then, the real-time PCR
results were compared. After leishmaniosis was found to be positive by real-time PCR,
species identification was performed using the HRMA method.
In microscopic examination, Leishmania amastigotes were seen in 27 (34.2%) of 79
patients with suspected CL, and they were considered positive. 1 (16.7%) of 6 patients
with suspected VL was considered positive. In real-time PCR, 25 (31.6%) of 79 patients
with suspected CL were considered positive. No 6 patients with suspected VL were
considered positive. When 25 patients with leishmaniosis, who were found to be
positive by Real-time PCR, were examined by the HRMA method, it was found that 21
(84%) were infected by L. tropica, 3 (12%) were infected by L. major and 1 (4%) was
infected by L. infantum.
As a result, it was concluded that direct microscopic examination and N.N.N. culture
medium are enough in the diagnosis of leishmaniosis, but the real-time PCR and
HRMA methods would be useful in determining the causative agent and the possible
drug resistance.
Key Words: Leishmania, Leishmaniosis, High Resolution Melting Analysis,
Genotyping, Real-Time PCR
The present work was supported by the Research Fund of Istanbul University. Project
No. 54948