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Öğe An alternative DNA extraction method for detection of Blastocystis spp. in human fecal samples(Academic Press Inc., 2018) Karasartova, Djursun; Güreser, Ayşe Semra; Ruh, Emrah; Türegün Atasoy, Buse; Çalgın, Mustafa Kerem; Taşçı, Leyla; Taylan Özkan, Hikmet AyşegülPolymerase chain reaction (PCR) is an effective technique for diagnosis of Blastocystis infection. Notably, DNA isolation procedure is extremely critical for the PCR step. In the present study, a recently described extraction procedure, named as the “sand method” was modified and adapted for isolation of Blastocystis DNA. To evaluate its efficacy, the current method and QIAamp DNA Stool Mini Kit (Qiagen) were applied to fresh human stool samples. Our results indicated that, the mean DNA concentrations obtained by the sand method and the commercial kit were 48 and 55 ng/?l, respectively. Also, no DNA inhibitors were detected in two methods. The sand method was capable of detecting 16 parasites per 50 mg feces. DNA samples extracted by both methods were subjected to PCR. Blastocystis spp. were detected in 11 (31.4%) of 35 samples, and perfect agreement (?: 1.000) was found between the PCR-sand method and PCR-commercial kit method. The samples that were detected positive by PCR-sand method were successfully sequenced, and Blastocystis subtypes (STs) were identified as ST3, ST2 and ST1. In conclusion, the present study indicates that the sand method provides a simple, rapid and inexpensive procedure for reliable extraction of Blastocystis DNA from stool samples. © 2018 Elsevier Inc.Öğe Anti-Pneumococcal Vaccine-Induced Cellular Immune Responses in Post-Traumatic Splenectomized Individuals(Springer New York LLC, 2017) Karasartova, Djursun; Gazi, Umut; Tosun, Özgür; Güreser, Ayşe Semra; Şahiner, İbrahim Tayfun; Dolapçı, Mete; Taylan Özkan, Hikmet AyşegülSplenectomy is associated with increased risk of overwhelming post-splenectomy infections despite proper anti-pneumococcal vaccination. As most studies concentrated on vaccination-induced humoral immunity, the cellular immune responses triggered in splenectomized patients are not yet well studied. The present study aims to investigate this area as it can contribute to the development of more effective vaccination strategies.Five healthy and 14 splenectomized patients were vaccinated with pneumococcal conjugate polysaccharide vaccine (PCV) followed by pneumococcal polysaccharide vaccine according to the guidelines established by Advisory Committee on Immunization Practices. PBMC samples collected 0, 8, and 12 weeks after PCV immunization were in vitro stimulated with PCV. Levels of lymphoproliferation, T-H cell differentiation, and cytokine release were assessed by carboxyfluorescein succinimidyl ester labeling, intracellular cytokine staining, and ELISA, respectively.While T(H)1-dominated immune response was detected in both groups, asplenic individuals generated significantly lower levels of T(H)1 cells following in vitro stimulation. Similarly, levels of IFN-gamma, IL-4, and IL-17 release and lymphoproliferation were significantly lower in asplenic patients.According to our data, splenectomy negatively influences the levels of PCV-induced lymphoproliferation, T(H)1 differentiation, and cytokine release. Besides, PCV failed to induce T(H)17-dominant immune response which is crucial for protection against extracellular pathogens.Öğe Bacterial and protozoal pathogens found in ticks collected from humans in Corum province of Turkey(Public Library of Science, 2018) Karasartova, Djursun; Güreser, Ayşe Semra; Gökçe, Tuncay; Çelebi, Bekir; Yapar, Derya; Keskin, Adem; Çelik, Selim; Ece, Yasemin; Erenler, Ali Kemal; Usluca, Selma; Mumcuoğlu, Kosta Yani; Taylan Özkan, Hikmet AyşegülBackground: Tick-borne diseases are increasing all over the word, including Turkey. The aim of this study was to determine the bacterial and protozoan vector-borne pathogens in ticks infesting humans in the Corum province of Turkey. Methodology/Principal findings: From March to November 2014 a total of 322 ticks were collected from patients who attended the local hospitals with tick bites. Ticks were screened by real time-PCR and PCR, and obtained amplicons were sequenced. The dedected tick was belonging to the genus Hyalomma, Haemaphysalis, Rhipicephalus, Dermacentor and Ixodes. A total of 17 microorganism species were identified in ticks. The most prevalent Rickettsia spp. were: R. aeschlimannii (19.5%), R. slovaca (4.5%), R. raoultii (2.2%), R. hoogstraalii (1.9%), R. sibirica subsp. mongolitimonae (1.2%), R. monacensis (0.31%), and Rickettsia spp. (1.2%). In addition, the following pathogens were identified: Borrelia afzelii (0.31%), Anaplasma spp. (0.31%), Ehrlichia spp. (0.93%), Babesia microti (0.93%), Babesia ovis (0.31%), Babesia occultans (3.4%), Theileria spp. (1.6%), Hepatozoon felis (0.31%), Hepatozoon canis (0.31%), and Hemolivia mauritanica (2.1%). All samples were negative for Francisella tularensis, Coxiella burnetii, Bartonella spp., Toxoplasma gondii and Leishmania spp. Conclusions/Significance: Ticks in Corum carry a large variety of human and zoonotic pathogens that were detected not only in known vectors, but showed a wider vector diversity. There is an increase in the prevalence of ticks infected with the spotted fever group and lymphangitis-associated rickettsiosis, while Ehrlichia spp. and Anaplasma spp. were reported for the first time from this region. B. microti was detected for the first time in Hyalomma marginatum infesting humans. The detection of B. occultans, B. ovis, Hepatozoon spp., Theileria spp. and Hemolivia mauritanica indicate the importance of these ticks as vectors of pathogens of veterinary importance, therefore patients with a tick infestation should be followed for a variety of pathogens with medical importance. © 2018 Karasartova et al.Öğe Bacterial and protozoan agents found in Hyalomma aegyptium (L., 1758) (Ixodida: Ixodidae) collected from Testudo graeca L., 1758 (Reptilia: Testudines) in Corum Province of Turkey(Elsevier Gmbh, 2020) Arslan Akveran, Gönül; Karasartova, Djursun; Keskin, Adem; Comba, Arzu; Çelebi, Bekir; Mumcuoğlu, Kosta Yani; Taylan Özkan, Hikmet AyşegülHyalomma aegyptium (L., 1758) (Ixodida: Ixodidae) is a hard tick and the main host for adults are Palearctic tortoises of the genus Testudo, while larvae and nymphs are less host-specific and nymphs also attach to humans. In the present study, a total of 261 H. aegyptium ticks were removed from 26 Testudo graeca L., 1758 in Corum Province of Turkey. The most prevalent pathogens identified molecularly in the ticks were Hemolivia mauritanica (51.9 %), followed by Rickettsia aeschlimannii (32.6 %), Ehrlichia spp. (30.2 %), and Bartonella bovis (0.8 %). All samples were negative for Coxiella burnetii, Francisella tularensis, Anaplasma phagocytophilum, Babesia spp., Hepatozoon spp. and Theileria spp. Overall, 97.4 % of the examined adult ticks and 26.3 % of nymphs were infected with at least one pathogen, while 40.9 % of all ticks were infected with only one pathogen, 27.4 % with two pathogens, and 9.9 % with three pathogens, concomitantly. Overall, 80.8 % of the examined blood smears of tortoises were H. mauritanica-positive, and the mean intensity of parasitemia was 4.8 % (1-21). As a conclusion, since the examined tortoises were sampled in gardens and vineyards close to human habitation, and as a rela-tively large percentage of them were infested with ticks carrying pathogenic agents affecting also humans, the importance of tortoises, their ticks and pathogens in terms of the public health should be farther examined.Öğe Basic problems in serological diagnosis of cystic echinococcosis(AVES, 2019) Güreser, Ayşe Semra; Karasartova, Djursun; Taylan Özkan, Hikmet AyşegülCystic echinococcosis (CE), which occurs in rural areas during most seasons, is an important public health problem in Turkey. Although the disease does not discriminate among age and gender, its occurrence is greater in women aged 30-50 years who reside in rural areas and are in frequent contact with animals (1, 2)Öğe Blastocystosis in post-traumatic splenectomized patients(Elsevier Ireland Ltd, 2016) Karasartova, Djursun; Güreser, Ayşe Semra; Zorlu, Musa; Türegün Atasoy, Buse; Taylan Özkan, Hikmet Ayşegül; Dolapçı, MeteBackground/aims The aim of this study was to determine the prevalence and significance of intestinal protozoa, specially Blastocystis spp., and to perform PCR-based subtype classification for understanding the importance of Blastocystis spp. in the pathogenesis of gastrointestinal disorders in post-traumatic splenectomized patients. Materials and methods A total of 60 stool samples were obtained from 30 post-traumatic splenectomized patients and 30 healthy controls. Wet mounts, trichrome and Kinyoun acid-fast stained slides were prepared from the stool specimens. PCR was used for detecting the presence of Giardia spp., Entamoeba spp., Dientamoeba fragilis, Cryptosporidium spp., Blastocystis spp. Genotyping was realized by using Blastocystis hominis STS primers. Results In both study groups, any helminth eggs and other protozoa except Blastocystis spp. were not detected by microscopy and PCR, and also bacterial cultures were negative. Only stool microscopy was positive for Blastocystis spp. in 30% (9 of 30) of splenectomized patients and in 13% (4/30) of healthy controls. PCR for Blastocystis spp. was positive in 40% (12 of 30), B. hominis genotypes were 20% (6/30): STS1 in 10% (3/30) and STS3 in 10% (3/30) of splenectomized patients. In healthy controls Blastocystis spp. was 13% (4/30) by PCR and genotypes of B. hominis was not detected. The difference between the prevalence of Blastocystis spp. infection in splenectomized patients and control groups was statistically significant (p = 0.020). Abdominal pain was the most frequent gastrointestinal symptom (p = 0.019) among splenectomized patients positive for Blastocystis spp. Conclusion In post-traumatic healthy splenectomized patients, Blastocystis spp. were found to be the most prevalent protozoa and may be responsible for the gastrointestinal disorders. © 2015 Elsevier Ireland LtdÖğe Çorum ilinde sokak köpeklerini enfeste eden kene türlerinin belirlenmesi(Refik Saydam National Public Health Agency (RSNPHA), 2020) Arslan Akveran, Gönül; Karasartova, Djursun; Comba, Arzu; Comba, Bahat; Keskin, Adem; Taylan Özkan, Hikmet AyşegülObjective: The Province of Çorum is an endemic area for tick-borne diseases such as the Crimean Congo hemorrhagic fever. The aim of this study was to identify the tick species infesting stray dogs found in the surrounding of human habitations. Methods: Hundred stray dogs kept in the Animal Nursing Home of the Veterinary Service of the province were randomly selected during the period of April 2018 and March 2019. Ticks were removed with the help of forceps and placed in 96% ethyl alcohol and stored at +4°C until they were taxonomically identified. Results: The following tick species were found: Ixodes kaiseri, Hyalomma spp., Rhipicephalus sanguineus, Rhipicephalus turanicus and Haemaphysalis parva. The infestation prevalence was 20%, the infestation density 3.3 (1-7) and the abundance was 0.7. The highest numbers of tick were recorded in June and August, while the lowest in April and May. Conclusion: The number of ticks collected from dogs in this study was relatively low, showing that the efforts of the Veterinary Services to control ticks infesting dogs, is successful. This should lower the possibilities of tick-borne diseases and zoonoses which could be transmitted by stray dogs in the region. © 2020. Turk Hijyen ve Deneysel Biyoloji Dergisi. All rights reserved.Öğe Çorum'da akut gastroenteritli çocuklarda rotavirüs ve adenovirüs saptanma sıklığı(Bilimsel Tıp Yayınevi, 2017) Güreser, Ayşe Semra; Karasartova, Djursun; Taşçı, Leyla; Boyacıoğlu, Zehra İlkay; Taylan Özkan, Hikmet AyşegülGiriş: Çocukluk çağı akut gastroenteritlerine en sık rotavirüs ve adenovirüs yol açmakta ve kesin tanı için laboratuvar testleri gerekmektedir. Çalışmamızda, hastanemize başvuran akut gastroenteritli çocuklarda immünokromatografik yöntemle saptanan rotavirüs ve adenovirüs sıklığının retrospektif olarak değerlendirilmesi amaçlanmıştır.Materyal ve Metod: Ocak 2013-Eylül 2014 tarihleri arasında, akut gastroenterit ön tanısıyla hastanemize başvuran 3189 çocuğa (0-18 yaş arası) ait dışkı örnekleri rotavirüs ve adenovirüs için kalitatif immünokromatografik kaset test (AbonBiopharm Rota/Adeno, Çin) ile çalışılmış, kayıtlar retrospektif olarak değerlendirilmiştir. İstatistiksel analiz ki-kare testi ile yapılmış, p< 0.05 olduğunda fark istatistiksel olarak anlamlı kabul edilmiştir.Bulgular: Hastaların 706 (%22.1)'sında viral antijen tespit edilmiştir. Dışkı örneklerinin %17.5'inde rotavirüs, %3.3'ünde adenovirüs, %1.3'ünde ise rotavirüs ve adenovirüs birlikte saptanmıştır. Rotavirüs ve adenovirüsün saptanma sıklığı açısından cinsiyetler arasında istatistiksel olarak fark olmadığı görülmüştür (p> 0.05). Rotavirüs antijeni en sık 7-24 ay (%22) ve 25 ay-4 yaş (%21.2) arasında, en nadir ise 5-10 yaş (%14.8) ve 11-17 yaş (%3.1) arasında pozitif olarak saptanmıştır (sırasıyla p< 0.0001, p= 0.042, p= 0.042 ve p< 0.0001). Sıfır-altı ay arasında ise %14.8 ile düşük saptanmış olmasına rağmen bulgumuz istatistiksel olarak anlamlı değildir (p> 0.005). Adenovirüs antijeni en sık 25 ay-4 yaş (%4.6), en nadir de 11-17 yaş (%0.1) arasında saptanmıştır (p= 0.007 ve p= 0.0006). Mevsimsel dağılım incelendiğinde ise rotavirüs en sık ilkbahar (%25.5), kış (%25.2) ve sonbahar (%23.5) mevsimlerinde, en az da (%5.9) yaz mevsiminde saptanmıştır (p< 0.005).Sonuç: Sonuç olarak bölgemizde özellikle Kasım ve Nisan ayları arasında akut gastroenterit şikayetiyle gelen 7 ay-4 yaş arasındaki çocuklarda rotavirüs ve adenovirüs etkenlerinin rutin olarak araştırılması gerekmektedir. Çocuklarda akut gastroenteritlerde viral etkenlerin saptanması hastalara gereksiz antibiyotik verilmesini önleyerek etkene yönelik tedavi uygulanmasına, çocukların ağır dehidratasyonlarının önüne geçilmesine ve aşılama çalışmalarının planlanmasına yardımcı olacaktır. Ayrıca, viral gastroenteritlere karşı koruyucu etkisi nedeniyle iki yaşına kadar bebeklerin anne sütü ile beslenmesi teşvik edilmelidir.Öğe Epidemiology and prevalence of blastocystis spp. in North Cyprus(American Society of Tropical Medicine and Hygiene, 2017) Seyer, Ayşe; Karasartova, Djursun; Ruh, Emrah; Güreser, Ayşe Semra; Turgal, Ebru; Imir, Turgut; Taylan Özkan, Hikmet AyşegülThis study was conducted to investigate the prevalence of Blastocystis spp. and its subtypes (STs) in North Cyprus; and to evaluate the presence of this parasite and its STs with respect to demographic, socioeconomic, and epidemiological factors, as well as gastrointestinal symptoms. Stool samples were collected from 230 volunteers. Each participant also filled out a questionnaire. The samples were examined microscopically by native-Lugol and trichrome methods and further tested by polymerase chain reaction (PCR) and sequencing. Prevalence of Blastocystis spp. infection was found to be 10.5%, 10.5%, and 27.8%, by direct microscopy, trichrome method, and PCR, respectively. No other parasites were detected in the specimens except Giardia spp. (n = 2; 0.8%) and Entamoeba coli (n = 1; 0.4%). The most common Blastocystis STs were ST3 (20; 31.2%), ST2 (18; 28.2%), ST1 (8; 12.5%), and ST4 (7; 11%); whereas other STs were identified as ST6 (3; 4.7%), ST7 (2; 3.2%), and non-ST (6; 9.4%). Presence of Blastocystis spp. and its STs was not significantly related to any of the demographic, socioeconomic, and epidemiological factors. Furthermore, no significant association of Blastocystis spp. and its STs with gastrointestinal symptoms was found. This study is the first investigation of the epidemiology of Blastocystis spp. in North Cyprus. Distribution of Blastocystis spp. and its STs among demographic, socioeconomic, and epidemiological factors showed complete homogeneity. Presence of the parasite and its STs was not significantly related with the gastrointestinal symptoms among symptomatic and asymptomatic individuals. These findings suggest that Blastocystis spp. may be part of the intestinal flora in humans. Copyright © 2017 by The American Society of Tropical Medicine and Hygiene.Öğe Hitit Üniversitesi Çorum Eğitim ve Araştırma Hastanesinde kan kültürlerinden üreyen mikroorganizmalar ve antimikrobiyal duyarlılıkları(Bilimsel Tıp Yayınevi, 2016) Taşçı, Leyla; Güreser, Ayşe Semra; Boyacıoğlu, Zehra İlkay; Karasartova, Djursun; Taylan Özkan, Hikmet AyşegülGiriş: Bakteremi ve sepsis tanısında altın standart kan kültürüdür. Kan kültürlerinde üreyen mikroorganizmaların dağılımı ve antibiyotik duyarlılıklarının belirlenmesi mortalite ve morbiditenin azaltılmasında çok önemlidir. Bu çalışmada laboratuvarımıza, hastanemizin yoğun bakım ünitesi ve diğer servislerinden gönderilen kan kültürlerinde üreyen mikroorganizmaların dağılımı, genişlemiş spektrumlu beta-laktamaz (GSBL) pozitifliği ve çeşitli antibiyotiklere duyarlılıklarının araştırılması amaçlanmıştır. Materyal ve Metod: Hitit Üniversitesi Çorum Eğitim ve Araştırma Hastanesi Mikrobiyoloji Laboratuvarında, 01 Ocak 2014-31 Mart 2015 tarihleri arasında 3100 hastadan alınan kan kültürleri BACT/ALERT 3D (bioMerieux, Fransa) otomatize kan kültür sisteminde inkübe edilmiştir. Sinyal alınan şişelerden kanlı agar, eozin metilen blue (EMB) ve çikolatamsı agar besiyerlerine ekim yapılmıştır. Plaklar 37°C'de 18-24 saat süre ile inkübe edilerek üreyen mikroorganizmaların (Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Staphylococcus aureus, Enterococcus spp.) VITEK 2 (bioMerieux, Fransa) tam otomatik bakteri tanımlama cihazı ile tür tayinleri ve antibiyotik duyarlılıkları belirlenmiştir.Bulgular: Kan kültüründe üreyen 224 suştan 80 (%35.7)'inde gram-pozitif, 144 (%64.3)'ünde gram-negatif bakteri saptanmıştır. GSBL pozitifliği E. coli suşlarında %37.2, K. pneumoniae suşlarında ise %29.4 oranında belirlenmiştir. Üreyen S. aureus suşlarının %25'inin metisiline dirençli olduğu görülmüştür. S. aureus ve Enterococcus spp. suşları tigesiklin, vankomisin ve teikoplanine yüksek oranda, Enterobacteriacealar ise karbapenem, kolistin ve aminoglikozidlere yüksek oranda duyarlı bulunmuştur.Sonuç: Kan kültürlerinde üreyen mikroorganizmaların izolasyonu, doğru tanımlanması ve antibiyotik duyarlılıklarının belirlenmesi ampirik tedavi uygulama sırasında yol göstericidir. GSBL üreten bakteriler pek çok antibiyotiğe dirençli oldukları için geniş spektrumlu antibiyotiklerin kullanılması gerekmektedir.Öğe Importance of NK Cells in Cellular and Humoral Responses Triggered by Pneumococcus Vaccination(KARGER, 2023) Gazi, Umut; Tosun, Özgür; Derici, Mehmet Kürşat; Karasartova, Djursun; Güreser, Ayşe Semra; Taylan Özkan, Hikmet AyşegülIntroduction: Despite the success of vaccination in reducing overall rate of pneumococcal pneumonia, Streptococcus pneumoniae is still held responsible for high mortality and modality rates worldwide. Our study aimed to investigate the potential role played by NK cells in immune response generated by pneumococcal vaccination, which could contribute to the development of more effective vaccines. Methods: The study included mice with and without NK cell depletion which were immunized with pneumococcus polysaccharide-conjugated vaccine followed by pneumococcus polysaccharide vaccine (PPV). Serum samples and splenocytes were collected from mice sacrificed 4 weeks after the last PPV dose. Serum samples were used for antibody level quantification by ELISA assay, while splenocytes were treated with PPV in vitro before monitoring CD4+ T-cell subsets (TH1, TH2, and TH17) and cytokine (IFN-?, IL-4, and IL-17) secretion levels by flow cytometry and ELISA analysis, respectively. Results: Results demonstrated reduced pneumococcal IgG and TH1 cell levels due to NK cell depletion. Nevertheless, in contrast to these observations, IFN-? secretion levels after in vitro PPV-23 treatment of splenocytes did not exhibit any statistically significant difference between the two mice groups. Conclusions: The data indicate a positive contribution of NK cells to both T-cell and B-cell responses triggered against pneumococcal vaccination. Further studies are required to confirm our data and investigate the potential benefit of NK cell targeting in promoting vaccine efficacy, especially in the elderly population who continues to be affected significantly by pneumococcal pneumonia.Öğe Influence of Maternal Toxoplasmosis on the Second-Trimester Aneuploidy Screening Test(2018) Görkem, Ümit; Güreser, Ayşe Semra; Toğrul, Cihan; Karasartova, Djursun; Güngör, Tayfun; Taylan Özkan, Hikmet Ayşegül; Koçak, ÖzgürOBJECTIVE: With apoptosis being critical for the development and homeostasis of placental tissues, it is possible to hypothesize that accelerated trophoblastic apoptosis during pregnancy may result in a partial loss of trophoblastic activity or trophoblastic cell mass, and ultimately may alter the second-trimester screening test parameters. Thus, this study was conducted to investigate the influence of maternal toxoplasmosis on second-trimester aneuploidy screening tests. STUDY DESIGN: This retrospective study was conducted with 552 pregnant women admitted to our University Hospital. The demographic data such as maternal age and weight; and the main parameters of second-trimester aneuploidy screening test including maternal serum alpha-fetoprotein, unconjugated estriol (uE3) and human chorionic gonadotropin were analyzed with the comparison of their Toxoplasma immunoglobulin serology results. RESULTS: The mean age of the pregnant women was 27.6 (17.0 - 43.0) years, and the mean maternal weight was 65.0 (40.0 - 120.0) kg. The pregnant women with positive Toxoplasma IgG antibody had a higher mean maternal age than those with negative Toxoplasma IgG antibody (p<0.0001). No significant difference for the concentrations and MoM values of second-trimester screening test parameters in women with Toxoplasma IgM and IgG antibodies was observed (p>0.05, for all). CONCLUSION: Although IgG-seropositivity of toxoplasmosis may lead to an accelerated trophoblastic apoptosis during pregnancy, there is no significant influence on the second-trimester screening test results. There was no data regarding the unaffected population whom used for calculation of MoM, if they had toxoplasmosis in their life span.Öğe Is “dried stool spots on filter paper method (DSSFP)” more sensitive and effective for detecting Blastocystis spp. and their subtypes by PCR and sequencing?(Springer Verlag, 2016) Seyer, Ayşe; Karasartova, Djursun; Ruh, Emrah; Güreser, Ayşe Semra; Imir, Turgut; Taylan Özkan, Hikmet AyşegülPCR and DNA sequencing are currently the diagnostic methods of choice for detection of Blastocystis spp. and their suptypes. Fresh or frozen stool samples have disadvantages in terms of several aspects such as transportation, storage, and existence of PCR inhibitors. Filter paper technology may provide a solution to these issues. The aim of the present study was to detect Blastocystis spp. and their subtypes by employing two different preservation methods: conventional frozen stool (FS) and dried stool spots on filter paper (DSSFP). Concentration and purity of DNA, sensitivity of PCR, and DNA sequencing results obtained from the two methods were also compared. A total of 230 fecal samples were included and separated into two parts: one part of the fecal samples were directly frozen and stored at ?20 °C. The remaining portion of the specimens were homogenized with saline and spread onto the filter papers as thin layer with a diameter of approximately 3 cm. After air-dried, the filter papers were stored at room temperature. DSSFP samples were collected by scraping from the filter papers. DNA were extracted by EURx Stool DNA Extraction Kit from both samples. Concentration and purity were measured with Nano-Drop, then PCR and sequencing were conducted for detection of Blastocystis spp. and its genotypes. Pure DNA was obtained with a A260/A280 ratio of 1.7–2.2 in both methods. DNA yield from FS was 25–405 ng/?l and average DNA concentration was 151 ng/?l, while these were 7–339 and 122 ng/?l for DSSFP, respectively. No PCR inhibition was observed in two methods. DNA from DSSFP were found to be stable and PCR were reproducible for at least 1 year. FS-PCR- and DSSFP-PCR-positive samples were 49 (21.3 %) and 58 (25.3 %), respectively (p = 0.078). The 43 specimens were concordantly positive by both FS-PCR and DSSFP-PCR. When the microscopy was taken as the gold standard, sensitivity of DSSFP-PCR and FS-PCR was 95.5 and 86.4 %, while specificity of both tests was 99.4 and 98.3 %, respectively. DNA sequencing results of 19 microscopically confirmed cases were strictly identical (concordance 100 %) in both methods, and ST2:6, ST3:8, ST4:3, and ST6:2 were the detected subtypes. Among the 230 fecal samples, the most predominant subtypes were ST3, ST2, ST4, and ST1 by both FS and DSSFP methods. Concordance of DNA sequencing results obtained from the two methods was noted to be 90.7 %. To our knowledge, this is the first study that demonstrates DNA extraction from DSSFP is more sensitive and effective than the FS method for diagnosis of Blastocystis spp. and their subtypes by PCR and DNA sequencing. © 2016, Springer-Verlag Berlin Heidelberg.Öğe Molecular Survey of Babesia microti (Aconoidasida: Piroplasmida) in Wild Rodents in Turkey(Oxford Univ Press Inc, 2019) Usluca, Selma; Çelebi, Bekir; Karasartova, Djursun; Güreser, Ayşe Semra; Matur, Ferhat; Öktem, M. Ali; Taylan Özkan, Hikmet AyşegülBabesia microti (Aconoidasida: Piroplasmida) (Franca, 1910) is an important tick-borne zoonotic parasite with rodents serving as reservoir hosts. In the present study, 536 rodents were captured from Burdur, Bartin, Giresun, and Yozgat provinces of Turkey between the years 2010 and 2012, and blood samples were examined for the presence of Babesia spp. using conventional PCR which targeted the 18S rRNA gene. The sequence analysis of PCR amplicons was tested for B. microti as well as for Hepatozoon spp., and Sarcocystis spp. Overall, 5.8% of the rodents were positive for B. microti: 41% in Myodes glareolus, 7.7% in Chionomys roberti, and 2% in Apodemus spp., whereas no Babesia DNA was detected in Mus macedonicus and Microtus spp. Six rodents were positive for Hepatozoon spp. and one rodent was positive for Sarcocystis spp. Overall, 14.9 and 4.5% of rodents captured from Bartin and Giresun provinces, respectively, were PCR positive for B. microti, whereas none of rodents captured in Burdur and Yozgat were positive for Babesia spp. The sequence data of B. microti from rodents revealed that all sequences belonged to the zoonotic genotype. Sequences of B. microti obtained from rodents of the Bartin province were genotypically closer to European isolates, whereas those obtained from rodents of the Giresun province were closer to Russian and Mongolian isolates.Öğe Nükleik asit ekstraksiyonunda kullanılmak üzere, gram-pozitif bakteriler ve mikobakterilerin hücre duvarlarının ortadan kaldırılmasında yeni bir yöntem: kum yöntemi(Ankara Mikrobiyoloji Derneği, 2016) Şahin, Fikret; Kıyan, Mehmet; Karasartova, Djursun; Çalgın, Mustafa Kerem; Akhter, Shameem; Türegün Atasoy, BuseGünümüzde moleküler yöntemler, enfeksiyon etkenlerinin hızlı tanısında yaygın olarak kullanılmaktadır. Bu amaçla en sık olarak polimeraz zincir reaksiyonu (PCR) yöntemi tercih edilmektedir. PCR kullanımında yeterli ve saf DNA veya RNA elde edilmesi önemlidir. Gram-negatif bakterilerde fenol-kloroform, DNAzol, proteinaz K, cam boncuk, kaynatma gibi farklı DNA ekstraksiyon yöntemleri başarıyla kullanılmaktadır. Hücre duvarında daha kalın peptidoglikan tabakası olan gram-pozitif bakteriler ile kompleks glikolipid bulunan mikobakteri cinsi bakterilerde ise, DNA ve RNA izolasyonu için bu kompleks yapıların ortadan kaldırılması gerekmektedir. Bu amaçla, stafi lokok cinsi gram-pozitif bakterilerde lizostafi n kullanılarak sferoplast oluşturulması, mikobakterilerde setil-trimetil amonyum bromür gibi kimyasallar kullanılarak bakteri duvarının tamamen veya kısmen uzaklaştırılması gerekmektedir. Herhangi bir kimyasal ajana gerek duyulmaksızın, bakteri hücre duvarının mekanik olarak ortadan kaldırılmasının amaçlandığı bu çalışmada, ince elenmiş kum partikülleri kullanılmış ve yöntem “kum yöntemi” olarak adlandırılmıştır. DNA ekstraksiyonu amacıyla ince elenmiş kum, küçük partiküllerini kaybetmeyecek şekilde ddH2O ile yıkanmış ve otoklavda sterilize edilmiştir. RNA ekstraksiyonu için ddH2O ile yıkanan kum, daha sonra %10 HCl’de (30 dakika) inkübe edilmiş ve otoklavda sterilize edilmiştir. Çalışmada, daha önce klinik bir örnekten izole edilen ve tanımlanan metisiline dirençli Staphylococcus aureus (MRSA) suşu, 100 mg hazırlanan kum ve 100 ?l Tris-EDTA çözeltisi içerisinde, 5 dakika maksimum hızda vortekslenmiş, DNA eldesi için proteinaz K ile muamele edilmiş ve sonrasında fenol kloroform-etanol presipitasyon protokolü takip edilmiştir. Kum yönteminin diğer yöntemler ile kıyaslanması amacıyla, kum yönteminde kullanılan miktardaki bakteri lizostafi n ile bir saat inkübe edildi ve kum yönteminde kullanılan miktarda proteinaz K eklenerek DNA ekstraksiyon işlemi tamamlanmıştır. Boncuk yönteminde steril cam boncuklar kum yönteminde olduğu şekilde kullanılmıştır. RNA eldesi için Mycobacterium tuberculosis H37Rv ATCC 25618, M.tuberculosis H37Ra ATCC 25177 ve M.tuberculosis H37Rv Pasteur Enstitüsü RSKK 598 standart suşları, 100 mg hazırlanan kum ve 20 ?l Tris-EDTA çözeltisi içerisinde 3-5 dakika maksimum hızda vortekslenmiş ve guanidinyum tiyosiyanat-fenol-kloroform (GTFK) ile klasik RNA ekstraksiyon protokolü tamamlanmıştır. Elde edilen DNA’nın kullanılabilirliği, çalışmada kullanılan MRSA suşundaki stafi lokinaz ve enterotoksin genlerine özgül primerler kullanılarak PCR ile araştırılmıştır. Kum yöntemi ile elde edilen RNA’nın kullanılabilirliğini saptamak için, önce cDNA sentezi yapılmış; daha sonra M.tuberculosis atım pompa genlerinden Rv1410c, Rv2333c ve DrrA’ya özgül primerler kullanılarak PCR uygulanmıştır. Karşılaştırma amacıyla, kum yönteminde kullanılan miktardaki mikobakteriye doğrudan GTFK protokolü uygulanmıştır. MRSA suşlarından lizostafi n uygulanarak elde edilen DNA ile kum yöntemi uygulanarak elde edilen DNA’lar agaroz jelde yürütülmüş, spektrofotometrede miktar ve saflıkları kıyaslanmıştır. Sonuç olarak, yaklaşık aynı miktar ve saflıkta DNA’ların elde edildiği gözlenmiştir. Kum yöntemiyle elde edilen DNA’ların herhangi bir inhibitör ajan içermediği, PCR’nin etkin olarak çalışmasıyla gösterilmiştir. Mikobakteri suşlarından kum yöntemiyle RNA elde edilebildiği halde, diğer yöntemlerle RNA elde edilememiştir. Kum yöntemi kullanılarak elde edilen RNA’ların gerek cDNA sentezinde gerekse bu cDNA’ların kullanıldığı PCR yönteminde etkin olarak çalıştığı gösterilmiştir. Çalışmamızda tanımlanan kum yöntemi ile nükleotid eldesi zor olan sert ve kompleks hücre duvarına sahip bakterilerden, saf ve yeterli miktarda DNA ve RNA elde edildiği belirlenmiştir. Sonuç olarak bu yöntemin, pahalı sayılabilen lizostafi n veya farklı kimyasalların yerine herhangi bir masrafı olmayan kumun kullanılması sayesinde ekstraksiyon maliyetini düşüreceği, ayrıca DNA ekstraksiyon süresini kısaltması açısından avantajlı olacağı düşünülmüştürÖğe Rozasea hastalarında demodex spp'nin araştırılması(Hitit Üniversitesi, 2019) Koşar, Nezahat; Sabancılar, Emine; Karasartova, Djursun; Güreser, Ayşe Semra; Öztekin, Aynure; Derici, M. Kürşat; Gazi, Umut; Artüz, Refika Ferda; Taylan Özkan, Hikmet Ayşegülİnsizyonel herni gelişiminde risk faktörlerinin saptanması Amaç: Rozasea, yüz bölgesinde çeşitli klinik bulgularla seyreden, değişik alt tipleri olan ve sebebi tam olarak bilinmeyen kronik enflamatuvar bir cilt hastalığıdır. Demodex spp. rozasea etyolojisinde rol oynadığı düşünülen insan derisinin zorunlu paraziti olan bir akardır. Çalışmanın amacı rozasea tanılı kadın hastalarda demodeks saptanma sıklığını ve rozasea alt tipleri ile parazitin saptanma yoğunluğu arasındaki ilişkiyi saptamaktır. Gereç ve Yöntem: T.C. Sağlık Bakanlığı Hitit Üniversitesi Erol Olçok Eğitim ve Araştırma Hastanesi Deri ve Zührevi Hastalıklar polikliniklerine Şubat-Aralık 2017 tarihleri arasında rozasea nedeniyle başvuran 27-73 yaş arasındaki 39 kadın hasta çalışmaya dahil edilmiştir. Hastalardan burun, yanak ve çene derisinden ayrı ayrı olmak üzere birer adet toplamda üç adet örnek standart yüzeyel deri biyopsisi yöntemi ile alınarak ışık mikroskobunda incelenmiştir. Bulgular: Otuz dokuz hastanın 34 (%87,2)'ünde demodeks saptanmıştır. Hastaların 14 (%35,9)'üne eritematotelenjiektatik; 25 (%64,1)'ine papülopüstüler rozasea tanısı konulmuştur. Eritematotelenjiektatik hasta grubundakilerin 11 (%78,5)'inde, papülopüstüler hasta grubundakilerin 23 (%92)'inde demodeks saptanmıştır. İki grup arasında yapılan istatistiksel analizde demodeks yoğunluğu ile rozasea alt tipi arasında anlamlı bir ilişki bulunamamıştır (p=0,276). Sonuç: Rozasea hastalarında demodeks enfestasyonun yüksek oranlarda görülmesi nedeniyle tanıda araştırılması gerektiği akıldan çıkarılmamalıdır. Ayrıca rozasea alt tiplerinde parazitin etkisini ve hastalığın etiyopatogenezini daha iyi ortaya koymak için daha fazla sayıda hasta grubu ile ilave çalışmalar yapılması faydalı olacaktır.Öğe Şanlıurfa yöresindeki Anofel larvalarının morfolojik tanımlanması ve üreme alanlarının fiziksel ve ekolojik özelliklerinin araştırılması(Refik Saydam National Public Health Agency (RSNPHA), 2020) Topluoğlu, Seher; Karasartova, Djursun; Karaer, Zafer Kadri; Taylan Özkan, Hikmet AyşegülObjective: Identification of vector species and determination of physical and ecological features of their breeding places is essential in implementation of scientific based mosquito control activities. In this study, it is aimed to identify Anopheles species by morphological method and determination of physical and ecological characteristics of their breeding places in Sanliurfa territory. Methods: Mosquito larvae were collected between September 29 and October 03, 2009 from determined 9 breeding places in Birecik, Eyyubiye, Haliliye, Harran, Siverek and Viransehir districts of Sanliurfa province where malaria cases had been reported and four instar larvae were identified morphologically according to keys of DuBose ve Curtin (1965), Merdivenci (1984) and Darsie and Samanidou-Vojadjoglou (1997). Essential ecological parameters of water in breeding places were measured. Temperature and dissolved oxygen were measured by using ExStik® DO600 (Extech Instruments-USA); pH and conductivity were measured by using Hanna Instruments 98129 pH / Conductivity /TDS Tester (Hanna Instruments-Germany) and salinity was measured using ExStik®II EC400 Conductivity/TDS / Salinity/Temperature Meter (Extech Instruments-USA). Results: Of the 274 four instar larvae collected, 231 (%84,3) were identified as An. sacharovi and 41 (%14,96) of them were identified as An. superpictus. Two (0,73%) samples identified as Anopheles genus, species discrimination could not be done. In %88,89 (n=8) of nine breeding places An. sacharovi and in %11,11 (n=1) of total breeding places An. superpictus found to be dominant species according to the morphological results. Malaria vector An. sacharovi detected in all breeding places which had different pH values, dissolved oxygen proportions, electrical conductivity, water temperature and salinity proportions with horizontal vegetation. The limits of tolerance for essential ecological parameters of species found to be as: pH-7,77-9,18 (mean 8,53); electrical conductivity-310-1100 (?S/cm) (mean 496,91); dissolved oxygen (mg/l)-1,64-13,06 (mean 9,67); temperature of water-20,3-25,8 °C (mean 23,46); salinity 0,15-0,55 ppt (mean 0,24). The limits of tolerance for essential ecological parameters of An. superpictus species in study area measured as: pH 8,48; electrical conductivity 710 ?S/cm; dissolved oxygen 8,91 mg/l; temperature of water 25,8 °C; salinity 0,35. In statistical analysis of physical and ecological characteristics of mosquito breeding places; no significant difference between pH values (p=0,189) was found between An. sacharovi and An. superpictus breeding places but significant dif¬ference have been found in water temperature (p= 0,0000001), electrical conductivity (p= 0,0000001), salinity (p= 0,0000001) and dissolved oxygen (p= 0,001) values. Conclusion: An. sacharovi is thought to be considered to be primary malaria vector in Sanliurfa Province as it can become the dominant species in malaria endemic areas and also in areas where transmission reoccur due to its ecological flexibility. In this context, vector control strategies in Sanliurfa should be revised and planning should be done according to the characteristics of the vector. © 2020 Refik Saydam National Public Health Agency (RSNPHA).Öğe SEROPREVALENCE OF RICKETTSIOSIS IN PATIENTS WITH CRIMEAN -CONGO HEMORRHAGIC FEVER PRELIMINARY DIAGNOSIS IN THE CASE OF CORUM PROVINCE TURKEY(Parlar Scientific Publications (P S P), 2021) Akdoğan, Özlem; Yapar, Derya; Gürel, Büşra; Karasartova, Djursun; Güreser, Ayşe Semra; Savcı, Ünsal; Taylan Özkan, Hikmet AyşegülPeople engaged in agriculture and animal husbandry living in endemic areas are at high risk of tick-transmitted infectious diseases. Both Crimean-Congo Hemorrhagic Fever (CCHF) and rickettsial diseases can be transmitted as a result of tick bites. We aimed to evaluate the patients preliminarily diagnosed with CCHF at our clinic in terms of CCHF and rickettsia seropositivity, epidemiologic features and to compare ELISA and IFAT for serodiagnosis of rickettsiosis. Between 2014-2017, 265 patients who were followed up with a preliminary CCHF diagnosis at the Infectious Diseases and Clinical Microbiology Department of Hitit University Comm Erol Olcok Training and Research Hospital, were included in this study. Rickettsia was analyzed by ELISA IgG and 1gM (Vircell, Rickettsia conorii ELISA IgG+IgM, Spain), IFA (Vircell, Rickettsia conorii IFA IgG, Granada, Spain) and also by in house-PCR. According to the laboratory results for CCHF and Rickettsia patients were divided into two groups: (i) CCHF positive (+), (ii) Rickettsia seropositive (Rickettsia+; ELISA/IFA IgM and/or IgG positive). Of the 265 patients, 179 (67.55%) were male, and the average age was 49.04 (age range 18-90) years. In our study, CCHF virus positivity was 51.3%, while Rickettsia IgG+IgM positivity was 24.9%. In the diagnosis of rickettsiosis IFA and ELISA showed 99.62% agreement, but no PCR positivity was found. In total, CCHF+(n=136), CCHF-(n=129), Rickettsia+(n=66), cases were evaluated. In total, 123 (90.44%) of the patients who were positive for CCHF and 55 (84.62%) of patients seropositive for Rickettsia had applied to the hospital from rural areas (p>0.05). In both group, most of the cases have tick bite history (77.2% in CCHF+ group and 59.1% in Rickettsia+ group). In the group in which both agents were found to be negative, this rate decreased to 38.2% (p 0.98). Rickettsia was found to be seropositive in 39 (28.7%) of the 136 patients with CCHF positivity. Rickettsia was seropositive in 27 (20.9%) of the 129 patients with CCHF negativity. Except one case with positive Rickettsia IgM, other 65 cases with IgG positive were not considered as acute rickettsiosis. The fact that we live in an area where CCHF and rickettsial diseases are endemic requires us to keep these diseases in mind constantly. Although IFAT is considered as the reference test for serological diagnosis of rickettsiosis, ELISA could be an alternative. Rickettsial disease, a deadly but treatable disease, should be especially considered in patients who apply with a history of acute fever in endemic areas.Öğe Skin-homing T-cell responses associated with Demodex infestation and rosacea(Wiley, 2019) Gazi, Umut; Güreser, Ayşe Semra; Öztekin, Aynure; Karasartova, Djursun; Koşar Acar, Nezahat; Derici, Mehmet Kürşat; Taylan Özkan, Hikmet AyşegülAims Our aim was to investigate the skin-homing T-cell immune responses triggered in patients with Demodex infestation and/or rosacea. Methods Collected whole blood samples were divided into four groups: control subjects; nonrosacea patients with Demodex infestation (Demodex group); papulopustular rosacea (PPR) patients without Demodex infestation (Rosacea group); and PPR patients with Demodex infestation (Rosacea/Demodex group). Following ex vivo activation, skin-homing CLA+CD4+ T-cell subset levels were monitored by flow cytometry. Results When compared with control subjects, among skin-homing CD4+ T-cell subsets analysed, Demodex patients had higher T(H)9 and T-reg cell levels; Rosacea subjects displayed elevated T(H)1 cell levels; and Rosacea/Demodex patients exhibited increased frequencies of T(H)9 and T(H)22 cells. In contrast to Rosacea subjects, Rosacea/Demodex group members displayed higher T(H)2 cell levels; and when compared with Demodex groups, they had higher T(H)1 and T(H)2 but lower T-reg cell levels. Demodex group members also exhibited higher T-reg but lower T(H)1 and T(H)22 levels than Rosacea/Demodex group subjects. Conclusions The skin-homing T-cell responses associated with Demodex infestation and rosacea formation seem to influence each other. The present as well as future studies could contribute to the development of effective treatment strategies for demodicosis and rosacea.Öğe Splenektomi, OPSI ve korunma stratejileri(Refik Saydam National Public Health Agency (RSNPHA), 2019) Gazi, Umut; Karasartova, Djursun; Güreser, Ayşe SemraOne of the most important functions of the spleen, which is the largest secondary immune system of the body, is to clear blood from foreign substances by initiating an immune response against antigens carried by blood. There are abundant amounts of lymphoid tissue and cells in spleen including macrophages attacking encapsulated microorganisms and B-cells responsible for early IgM production. In the absence of the spleen, rapid antibody production against a newly encountered antigen is impaired and the bacteria can multiply rapidly. Post-splenectomy infection (OPSI) is a highly mortal disease. Although the initial symptoms of OPSI follow a mild course as in flu-like illnesses, the clinical course can quickly lead to coma and death within two days. Splenectomized patients are susceptible to develop OPSI and possess the risk for lifetime. OPSI cases are mostly caused by Streptococcus pneumoniae (S. pneumoniae), Neisseria meningitis (N. meningitis) and Haemophilus influenzae (H. influenzae). Therefore, pneumococcal, meningococcal and Hib vaccination is recommended at least two weeks prior to splenectomy treatment, or at most two weeks after surgery if emergency splenectomy is required. Since the antibody levels decrease in individuals vaccinated over time, splenectomy patients should be re-vaccinated for every 5 years. On the other hand, antibiotic use and patient training have also an important role in the prevention of OPSI. Patients need to be informed about the OPSI risk, and need to consult their doctor before going abroad. Even though it is so important the level of patient and physician education is not required to reduce OPSI risk today. Therefore, physicians are recommended to be more active in informating their patients abbout OPSI and carefully follow up their patients. In this review, the efficacy of the strategies used to prevent OPSI will be discussed in the context of current literature, including our own laboratory and clinical study results.
