Browsing by Author "Zeyrek, FY"
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Item Cutaneous leishmaniasis caused by Leishmania infantum in Turkey: reports of two cases diagnosed with genotyping and protein fingerprintingCulha, G; Akyar, I; Zeyrek, FY; Gündüz, C; Kurt, Ö; Ostan, I; Töz, S; Kocagoz, T; Ozbel, Y; Ozbilgin, AItem Cutaneous and visceral tropisms of Leishmania tropica/Leishmania infantum hybrids in a murine model: First report of hybrid Leishmania strains isolated in TurkeyOzbilgin, A; Culha, G; Zeyrek, FY; Töz, S; Gündüz, C; Kurt, Ö; Pratlong, F; Ozbel, YItem Evaluation of 2015-2016 MOTAKK HBV DNA and HCV RNA External Quality Assessment National Program ResultsKaratayli, E; Soydemir, E; Aksoy, ZB; Kizilpinar, M; Altay Koçak, A; Karatayli, SC; Yurdcu, E; Yildirim, U; Güriz, H; Bozdayi, G; Yurdaydin, C; Ilhan, O; Yildirim, Y; Bozdayi, AM; Oguz, AY; Baris, A; Alp, A; Aksözek, A; Sayiner, A; Karagul, A; Ordu, A; Istanbullu, A; Otlu, B; Aridogan, B; Aksu, B; Buruk, CK; Karahan, C; Güney, Ç; Toksöz, D; Yildirim, D; Çolak, D; Daglar, DE; Findik, D; Kas, E; Çaliskan, E; Zeyrek, FY; Arslan, F; Demir, F; Milletli, F; Kibar, F; Özdinçer, F; Dündar, G; Arslan, H; Agca, H; Aliskan, HE; Güdücüoglu, H; Fidan, I; Akyar, I; Afsar, I; Kaleli, I; Dönmez, I; Yanik, K; Midilli, K; Çubukçu, K; Özdemir, M; Acar, M; Yalinay, M; Kuskucu, MA; Bakici, MZ; Aydin, N; Yilmaz, N; Çeken, N; Ziyade, N; Yilmaz, N; Özgümüs, OB; Gitmisoglu, Ö; Demirgan, R; Kesli, R; Güçkan, R; Sertöz, R; Akgün, S; Aksaray, S; Tezcan, S; Kaygusuz, S; Gökahmetoglu, S; Mese, S; Bayik, SA; Akçali, S; Gürcan, S; Karsligil, T; Us, T; Özekinci, T; Pilgir, T; Aslan, U; Dinç, U; Coskun, USS; Çetinkol, Y; Keskin, Y; Ayaydin, Z; Toraman, ZAMOTAKK, as a national external quality control program has been launched to evaluate the molecular detection of viral infections including HBV DNA and HCV RNA in molecular microbiology diagnostic laboratories in Turkey. This program is prepared in compliance with ISO 17043:2010 (Conformity assessment general requirements for proficiency testing) standards, and aims to take the place of external quality control programs from abroad, contributing to standardization and accuracy of molecular diagnostic tests in our country. The aim of this study was to evaluate 2015 and 2016 results of the MOTAKK External Quality Control Program for HBV DNA and HCV RNA viral load. The calls were announced on the web page of MOTAKK (www.motakk.org). The quality control samples were sent to participating laboratories in 2015 and 2016. Main stocks were prepared from patients with chronic hepatitis B and C who had viral load detection with reference methods according to WHO reference materials for viral load studies to improve quality control sera. From these main stocks, samples with different viral loads were prepared from dilutions of plasma with HBV, HCV, HAV, HIV, Parvovirus B19 and CMV negative serologic markers. Quality control samples were sent to the participating laboratories along with the negative samples in the cold chain. The laboratories accomplished the related tests within 2-3 weeks and entered their results on the MOTAKK web page. These results were analysed according to ISO 13528 (Statistical methods for use in proficiency testing by interlaboratory comparison) and scoring reports were created by a software developed by MOTAKK and sent to participating labs. Each laboratory evaluated their own results in comparison with the other laboratory results, reassessed the tests via observing the distance from the mean result and the reference values. The number of laboratories participating in the HBV DNA and HCV RNA external quality control program was 70-73 in 2015-2016. Participants were able to comply with the program tools, registering, entering results and receiving the results reports problem. In HBV panel, 72.6-89.1% and 84.7-90.3% of the participant laboratories were in 1 standard deviation (SD) in 2015-2016, respectively. In HCV panel, 70.8-89.1% and 84.7-90.3% of the participant laboratories were in 1 SD in 2015-2016, respectively. A national external quality control program for HBV DNA and HCV RNA in Turkey has been prepared for the first time with this project and implemented successfully. All the data provided in the MOTAKK external quality control program final report, compensate all the data provided by the quality control program final reports from abroad; additionally, the report allows comparison of used technologies and commercial products.Item Determination of Antimony Resistance Mechanism of Leishmania tropica Causing Cutaneous Leishmaniasis in TurkeyÖzbilgin, A; Zeyrek, FY; Güray, MZ; Çulha, G; Akyar, I; Harman, M; Özbel, Y; Ertabaklar, H; Çavus, I; Gündüz, CWorld Health Organization reported that approximately one billion people are at risk in endemic areas, one million cases of cutaneous leishmaniasis (CL) and approximately 300,000 cases of visceral leishmaniasis (VL) were reported per year in the last five years. The number of deaths due to VL is reported to be approximately 20,000 per year. Approximately 2500 cases/year have been reported as CL, caused by Leishmania tropica and Leishmania infantum, in Turkey. The significant increase observed in many cities mainly in the provinces of Mediterranean and Aegean regions in cases and foci in recent years, suggests that there may be an increase in this infections in the following years as well. In Turkey, the causative agent of CL is L.tropica and meglumine antimoniate is used in the treatment of CL. We aimed to determine antimony resistance genes specific for L.tropica by comparing the gene and protein expressions of antimony-resistant and non-resistant L.tropica strains. Ltropica isolates obtained from 3 CL patients without antimonate resistance from Aegean, Mediterranean and Southeastern regions of Turkey were provided to transform into 3 resistant isolates against meglumine antimony in the laboratory conditions. Gene expression alterations by microarray method; protein profiles by two-dimensional gel electrophoresis (2D-PAGE) and relevant proteins by MALDI-TOF/TOF MS of these isolates were accomplished and compared. L.tropica isolates from 10 CL patients who did not respond to antimony therapy were analyzed for resistance to antimonial compounds and quantitative real-time polymerase chain reaction was performed to detect the expression of genes responsible for resistance development. Moreover, differences in protein expression levels in isolates with and without antimony resistance were determined by comparing protein profiles and identification of proteins with different expression levels was carried out. Enolase, elongation factor-2, heat shock protein 70, tripanthione reductase, protein kinase C and metallo-peptidase proteins have been shown to play roles in L.tropica isolates developing resistance to antimonial compounds and similar expression changes have also been demonstrated in naturally resistant isolates from patients. In conclusion, it was revealed that L.tropica strains in our country may gain resistance to meglumine antimoniate in a short time. It is foreseen that if the patients living in our country or entering the country are treated inadequately and incompletely, there may be new, resistant leishmaniasis foci that may increase the number of resistant strains and cases rapidly.Item Diversity of Leishmania Strains Isolated from Cutaneous Leishmaniasis Patients in Turkey and its Reflection to Clinics in Mice ModelÖzbilgin, A; Çulha, G; Güray, MZ; Zeyrek, FY; Akyar, I; Töz, S; Ural, IÖ; Kurt, O; Kocagöz, T; Çavus, I; Gündüz, CAlthough asexual reproduction has been attributed to Leishmania species, genetic exchange has recently been demonstrated, which helped emerging of hybrid isolates. Situated on the crossroads between three continents, Leishmania hybrids may be present in Turkey. In Turkey, visceral leishmaniasis caused by Leishmania infantum is less common, while cutaneous leishmaniasis (CL) caused by Leishmania tropica and L.infantum could reach 2500 reported cases a year. Our aim was to investigate genetic variability of local Leishmania species and presence of hybrid Leishmania strains in Turkey. Twenty CL patients from Sanliurfa and Hatay, where only L.tropica and both L.tropica and L.infantum cause CL, respectively, were registered equally. All isolates were assessed with real-time polymerase chain reaction (Rt-PCR), isoenzyme analysis, gene sequencing, two-dimensional gel electrophoresis (2D-PAGE) and MALDI-TOF/TOF-MS followed by in vivo analyses on mouse model. Identification of differentially expressed proteins was performed. These proteins were confirmed by sequence analysis. All isolates from Sanliurfa were found to be L.tropica which caused cutaneous infection in mice. However, one of 10 isolates from Hatay was found as Leishmania major which caused cutaneous infection. Five isolates were found as L.tropica with Rt-PCR and gene sequencing, one of which had one different protein from the reference L.tropica strain and caused cutaneous infection. Four of the five isolates had five different proteins compared to reference strain and caused both cutaneous and visceral infections. Remaining four isolates showed double melting curves in Rt-PCR, which were concordant with L.tropica and L.infantum. Their sequencing and isoenzyme analyses indicated them as L.infantum. They had six different proteins compared to reference L.infantum strain and caused cutaneous and visceral infections. It is concluded that the isolates with different proteins were hybrid Leishmania species. In the present study, outcomes of the proteomics, genomics, clinical manifestations and tissue tropism on animal models were evaluated together for the first time. In addition to L. tropica and L.infantum, L.major was identified as a causative agent for CL and hybrids of Linfantum/tropica were also shown to be present.Item Leishmaniasis in Turkey: Determination of Leishmania Species by Matrix-Assisted Laser Desorption Ionization Time-Of-Flight Mass Spectrometry (MALDI-TOF MS)Culha, G; Akyar, I; Zeyrek, FY; Kurt, Ö; Gündüz, C; Töz, SÖ; Östan, I; Cavus, I; Gülkan, B; Kocagöz, T; Özbel, Y; Özbilgin, ABackground: Cutaneous leishmaniasis (CL) is endemic in Southeastern Anatolia, mainly in Sanliurfa and Hatay provinces, and the causative agents are mostly Leishmania tropica and less frequently L. infantum. Here, we report the first MALDI-TOF analyses of Leishmania promastigotes obtained from the cultures of two CL cases from Osmaniye and Hatay provinces who were initially diagnosed by microscopy, culture and identified as L. infantum with Real-Time PCR (RT-PCR). Methods: Samples obtained from the skin lesions of patients were initially stained with Giemsa and cultivated in NNN medium. Examination of the smears and cultures revealed Leishmania amastigotes and promastigotes, respectively. The promastigotes (MHOM/TR/2012/CBU15 and MHOM/TR/2012/MK05) obtained from the cultures of both patients were used for RT-PCR targeting the ITS-1 region in the SSU of rRNA. The reference strains of four Leishmania species (L. infantum, L. donovani, L. tropica and L. major) were initially assessed with MALDI-TOF and their data were added to MALDI-TOF Biotyper Library. Results: Both RT-PCR and MALDI-TOF analyses indicated that the causative agent in both patient samples was L. infantum. Conclusion: Despite disadvantages such as requirement of culture fluid with nothing but promastigotes and high cost, MALDI-TOF analysis may be a fast, sensitive and specific diagnostic tool in especially large-scale research studies, where the cost declines, relatively.