Investigation of Sensitivity of Rapid Diagnosis Tests in Patients with Suspected Malaria

Objective: Malaria has been eradicated in Türkiye as of 2010, but there are imported cases. In this study, we aimed to compare the diagnostic value of two rapid tests; SD Bioline Malaria Ag Pf/Pan (SD-Pf/Pan) and SD Bioline Malaria Ag Pf/Pv (SD-Pf/Pv) with microscopy and real time-polymerase chain reaction (RT-PCR). Methods: Blood samples were taken from all participants. Thick drop smears were prepared. Thick drop smears were examined for malaria positive/negative distinction under the light microscopy. Then, two rapid diagnostic tests (SD-Pf/Pan and SD-Pf/Pv) were performed. After DNA extraction from blood samples, RT-PCR was typed. The data were evaluated with SPSS 21 program of statistics. Results: A total of 30 cases out of 66 suspected malaria cases were detected as positive with microscopy and RT-PCR. Twenty-seven patients were found positive with both SD-Pf/Pan and SD-Pf/Pv tests. Based on the microscopic results as a reference method, SD-Pf/Pan and SD-Pf/Pv rapid diagnostic tests had a 90% sensitivity, 100% specificity, 100% positive predictive value (PPV), and 92.86% negative predictive value (NPV). Based on the RT-PCR results as a reference method, for detection of P. falciparum, both tests had a 95.65% sensitivity, 100% specificity, 100% PPV, and 88.89% NPV. Moreover, while SD-Pf/Pv had a sensitivity, specificity, PPV, and NPV of 100% in detection of P. vivax; SD-Pf/Pan has a 77.78% sensitivity of, 61.90% specificity of, 46.67% PPV, and 86.67% NPV SD-Pf/Pan for detection of PAN. Conclusion: As a result, high sensitivity and specificity were detected in both kits in the diagnosis of malaria infections caused by P. falciparum and P. vivax. Rapid diagnostic tests can be used safely in diagnosis however the diagnosis should be supported by microscopy and RT-PCR methods when they are applicable.

Evaluation of Four Adult Visceral Leishmaniasis Cases.

Leishmania infantum is the species responsible for visceral leishmaniasis [(VL), kala-azar], which is observed sporadically mainly in pediatric age groups in the Aegean, Mediterranean and Central Anatolian regions of Türkiye. The aim of this study is to evaluate the diagnosis, clinic, laboratory results and treatments of four adult patients with VL who applied to our hospital. The patients were referred to our hospital to investigate hematological malignancy. In the study, the data of four patients (three men, one woman; age range: 30-40 years) who were diagnosed with VL and treated in the infectious diseases clinic of our hospital between January 2022 and April 2022 were evaluated retrospectively. The diagnosis of VL was made according to appropriate clinical and physical examination findings, biochemical and serological tests (indirect fluorescent antibody test and rK39 rapid antigen test) and polymerase chain reaction (PCR) results, as well as the presence of amastigote forms of the parasite in bone marrow samples. Serology positivity was found in all patients, and bone marrow positivity was found in two patients. According to the results of RT-PCR in all patients, it was determined that the species causing the disease was L. infantum/L. donovani. Initially, the most common symptoms were fever, fatigue, and abdominal distension. None of the patients had an immunosuppressive condition. It was understood that all the patients lived in the rural area of Syria's Idlib province. Hepatosplenomegaly, increased erythrocyte sedimentation rate, anemia, leukopenia and thrombocytopenia were found in all patients. The patients were treated with liposomal amphotericin-B (L-AMB). One patient did not come for follow-ups, the other three patients were found to have completely recovered in their follow-up. No recurrence was observed in any of the patients. In conclusion, VL should be considered in patients who apply to health institutions with complaints of fever, hepatosplenomegaly, increased erythrocyte sedimentation rate, anemia, leukopenia and thrombocytopenia.

A Case of Leishmania Infantum Mimicking Lymphoma.

Leishmaniasis is a parasitic infection caused by an obligate intracellular protozoon transmitted by infected sand flies. Cutaneous leishmaniasis (CL) is a skin disease known in our country as oriental sore, which heals, leaving a scar in place, mainly on the skin and sometimes in the mucous membrane. Demonstration of the parasite in chronic CL is difficult. Moreover, differential diagnosis from other granulomatous dermatitides such as lupus vulgaris, sarcoidosis and deep mycosis is growing difficult. A case of CL was presented in an 84-year-old female patient who had a pre-diagnosis of lymphoma and a nodule lesion on her forehead for 2.5 months. In the smear of the sample taken from the lesion, amastigote forms of the parasite were diagnosed and typed as L. infantum by the Real-Time Polymerase Chain Reaction (RT-PCR) method.

Cutaneous Leishmaniasis with Mucosal Involvement

Leishmaniasis is a protozoan parasitic disease transmitted to humans by infected female sand flies. Turkey has received more than three million immigrants from Syria because of the civil war and political instability. This study reported cases of two patients, who were from Syria and lived in Hatay, with cutaneous leishmaniasis and mucosal involvement. Two patients presented to the infectious diseases clinic with a complaint of facial lesions and were subsequently referred to the parasitology department laboratory. Smears were prepared from the lesions, stained with Giemsa and examined under a microscope. Moreover, aspirates taken from the patients' lesions were inoculated into the modified Novy-MacNeal-Nicolle medium. The diagnosis was made when amastigotes were detected in both smears. Proliferation of promastigotes was observed in one of the clinical specimens inoculated on the medium. By PZR-RFLP, Leishmania tropica were detected in the isolate. Both patients were treated with amphotericin B. One patient was treated again with a pentavalent antimony compound because of the recurrence of the lesion.

The amoebicidal activity of three substances derived from benzothiazole on Acanthamoeba castellanii cysts and trophozoites and its cytotoxic potentials.

Acanthamoeba species are free-living amoebae isolated from many ecological areas such as swimming pools, dams, lakes, soil, and air filters. These amoebae are usually causing granulomatous amebic encephalitis and amebic keratitis in immunosuppressive individuals. In this study, the reproductive potential and morphological changes determined of Acanthamoeba castellanii trophozoite and cyst forms exposed to three different active substances derived from benzothiazole. Furthermore, the cytotoxic potential of these active substances determined by XTT analysis. In the study, axenic cultures prepared for Acanthamoeba castellanii cyst and trophozoite forms and parasite exposed to different concentrations of active substances. Cell counts of parasite cultures were performed at the 30 minutes, 1st, 6th, 12th, 24th, and 48th hour periods. As a result of the study, the reproductive potential suppressive effects of all three substances on Acanthamoeba castellanii trophozoites and cysts were determined. The most effective of these substances was 2-Amino-6(trifluoromethoxy)-benzothiazole. In the first three concentrations of this substance (0.1%, 0.05%, 0.025%), no determined trophozoite and cysts at the end of twenty four. Due to its strong ameobicidal effect, it is thought that 2-Amino-6(trifluoromethoxy)-benzothiazole may be a new therapeutic agent in diseases caused by acanthamoeba parasites by supporting this study with animal experiments.

The Investigation of the Association of Cutaneous Leishmaniasis in Biopsy Specimens of the Patients with Granulomatous Disease and Skin Cancer Using the Molecular Method.

BACKGROUND: Clinically, cutaneous leishmaniasis (CL) can be confused with granulomatous diseases and skin cancers, and it may lead to erroneous diagnosis and treatment. Diagnosis based and histopathology can have some difficulties due to low number of parasites, especially in chronic CL cases. We aimed to emphasize the necessity of considering CL in the differential diagnosis for cases of granulomatous diseases and basal cell carcinoma, particularly in areas where CL is endemic. METHODS: One hundred and seven paraffin-embedded tissue biopsy specimens were selected from the archive, as of 2002, of Pathology Department, School of Medicine, University of Hatay Mustafa Kemal in Hatay, Turkey. After DNA isolation, performed with the samples were used for PCR analysis with specific 13A, 13B primers targeting kinetoplastid DNA (kDNA) found in all Leishmania species. Another PCR was performed with LITSR and L5.8S primers targeting ITS-1 internal-transcribed-spacer-1 (ITS-1) region to subtype positive samples. Then these samples were further analyzed for subtyping with PCR-RFLP using HaeIII enzyme (BsuRI). RESULTS: Ten out of 107 tissue specimens were positive via kDNA-PCR. Lupus vulgaris, sarcoidosis, skin lymphoma and Leishmania cutis appeared in 9 out of 10 positive specimens. One of the cases presented with a mass on the cheek and was pre-diagnosed with hemangioma, but leishmaniasis did not appear. All of 10 specimens were diagnosed as granulomatous dermatitis. Two out of 10 samples, found positive with kDNA-PCR, were analyzed with ITS-1-PCR and identified as L. infantum/donovani after RFLP. CONCLUSION: Molecular methods should be utilized in the differential diagnosis of CL to eliminate false diagnoses of granulomatous diseases and skin cancers.

[Diversity of Leishmania Strains Isolated from Cutaneous Leishmaniasis Patients in Turkey and its Reflection to Clinics in Mice Model]. / Türkiye'de Kutanöz Leysmanyazis Hastalarindan Elde Edilen Leishmania Izolatlarindaki Farkliliklar ve Bunlarin Fare Modeline Klinik Yansimasi.

Although 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/TOFMS 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 L.infantum/tropica were also shown to be present.

[Determination of Antimony Resistance Mechanism of Leishmania tropica Causing Cutaneous Leishmaniasis in Turkey]. / Türkiye'de Kutanöz Leysmanyazis Etkeni Leishmania tropica'da Antimon Direnç Mekanizmasinin Belirlenmesi.

World 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. L.tropica 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.

Genotyping of Cutaneous Leishmaniasis Cases Detected Before and After Migration with Real-Time Polymerase Chain Reaction in Hatay

Objective: Leishmania tropica (L. tropica) and Leishmania infantum (L. infantum) are the species causing cutaneous Leishmaniasis (CL) in Turkey. There was a wave of immigration due civil war in Syria in 2011. Migration from Syria, where CL is endemic, to other countries is thought to affect the number of CL cases and species diversity. The aim of the study was to typify the samples of CL positive, pre-migration and post-migration Turkish patients and importe (Syrian) patients whose smears were found in the archive and to reveal the difference of CL species before and after migration in Hatay. Methods: Smears of a total of 150 patients (50 Turkish patients before migration, 50 Turkish patients after migration and 50 Syrian patients) which had been prepared with dermal scraping, stained with Giemsa and determined as CL positive by microscope examination were included in the study. DNA isolation of selected preparations was performed and GZ-PZR analysis with ITS-1probe was performed for species determination. Results: L. infantum/donovani was detected in 40 (80%), L. tropica in 8 (16%), and L. major in 2 (4%) of the samples belonging to pre-immigration Turkish patients. L. infantum/donovani was detected in 28 (56%), L. major in 3 (6%) and L. tropica in 19 (%38) of the samples belonging to post-immigration Turkish patients. L. infantum/donovani was detected in 2 (4%), L. major in 1 (2%) and L. tropica in 47 (94%) of the samples belonging to Syrian patients. Conclusions: It was observed that in local cases in Hatay before immigration, L. infantum/donovani was the common species that caused CL and that after immigration L. tropica began to raise and that L. major was more encountered than before. It was concluded that Syrians coming to Hatay may have caused diversity in the Leishmania species which were the causative agents of CL, and that further research was needed on the subject.

[Investigation of Polymerase Chain Reaction Method in Patients with Suspected Chronic Cutaneous Leishmania of Negative Microscopy]. / Mikroskop Incelemesi Negatif Olan Süpheli Kronik Kutanöz Leishmania Olgularinin Polimeraz Zincir Reaksiyonu Yöntemi ile Arastirilmasi.

Leishmaniasis is a parasitic disease that is transmitted to humans by the bites of infected female phlebotomine sandflies. In the diagnosis of cutaneous leishmaniasis (CL), in the smear samples, the demonstration of the parasite by microscope remains a gold standard method. However, it becomes difficult to diagnose the parasite since the number of amastigotes in chronic cases with a lesion of one year or longer is very low. Due to many factor such as patients primarily do not to take any notice these lesions in their bodies, do not apply to health institutions or late applied, receive wrong treatment; the diagnosis and treatment are delayed. In addition, it is been worse prognosis by add secondary infection to lesions and wounds become chronic. For this reason, molecular methods are used in addition to microscopic examination in chronic suspected CL cases. It was aimed to reveal of the molecular diagnostic value in chronic suspected CL cases by polymerase chain reaction (PCR) in the smear belonging to Turkish patients that reported to be evaluated clinically because it can not be seen Leishmania amastigotes in microscopic examination. Smear of 50 Turkish patients who were clinically reported of the evaluation of chronic CL were selected. These samples were smears belonging to suspected CL patients that applied Hatay Mustafa Kemal University, Faculty of Medicine, Parasitology laboratory from different polyclinics and were decided to be evaluated clinically as a result of microscopic examination because they came from endemic regions (such as Hassa, Altinözü, Yayladagi). DNA was isolated from selected samples and PCR was performed using 13A, 13B primers targeting the kinetoplastid DNA (kDNA) region. The samples found positive by PCR were typed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis using LITSR and L5.8S primers targeting internal transcribed spacer (ITS-1) region. Of the 50 smear samples, 17 (34%) were determined positive with 13A, 13B primers targeting the kinetoplastid DNA (kDNA) region. Positive samples were also found to be positive with LITSR and L5.8S primers targeting ITS-1 region. The PCR products obtained from PCR with ITS-1 gene region were digested with the restriction endonucleases BsuRI (HaeIII). As a result of PCR-RFLP analysis, it was determined that 11 of Leishmania tropica, one of Leishmania major and five of Leishmania infantum/donovani out of 17 samples. Chronic CL can be confused with skin diseases such as sarcoidosis, tuberculosis, malignant tumors. In particular, chronic CL cases can be escaped the attention for many reasons such as failure to diagnose correctly, insufficient microscope experience, fail to see due to low number of parasites. For this reason, it was concluded that PCR, which is a molecular method, should be used in chronic suspected CL samples which are negative for the parasite by microscopic examination.

Retrospective Analysis of Cases with Imported Malaria in Hatay Province of Turkey: Seventy-Five Cases in Ten Years

Objective: Cases with imported malaria have increased complication and mortality rates because of delayed diagnosis and treatment in non-endemic countries. This study aimed to investigate the incidence and clinical features of imported malaria in our clinic during the past 10 years. Methods: This retrospective study included 75 cases diagnosed as having imported malaria in our clinic between January 2008 and December 2017. The epidemiological data, laboratory findings, treatment data and clinical course of the cases were obtained from system records. Results: Patients were predominantly male (%98.6) with a median age of 51 (23-64) years. All cases were infected with Plasmodium falciparum, had a recent travel history to Sub-Saharan African countries and none had received chemoprophylaxis before travel. The incidence of imported malaria showed a declining trend after 2015. The most common findings were fever (100%), thrombocytopenia (84%) and anemia (72%). Although 8% of patients had presented with severe malaria, none of them died. Conclusion: Despite increasing incidence of imported malaria in our country in recent years, there is a decrease in this number in our region. Since Turkey is one of the countries with the highest prevalence of imported malaria in the world, patients with fever and thrombocytopenia should be questioned whether or not they had a history of travel to malaria-endemic area.

[Imported cutaneous leishmaniasis cases detected in truck drivers in Hatay]. / Hatay'da tir soförlerinde saptanan yurt disi kaynakli kutanöz leysmanyazis olgular.

Leishmaniasis, seen in tropical and subtropical regions, is an infectious disease caused by the protozoan parasite Leishmania species. There are three main forms of leishmaniasis: cutaneous, mucocutaneous and visceral leishmaniasis. Cutaneous leishmaniasis (CL) has become an increasing problem as the number of travels around the world increases and people go to work in endemic areas. Turkey has received great numbers of immigrants in recent years, from its neighboring countries like Iraq, Islamic Republic of Iran, Afghanistan, Turkmenistan and the Syrian Arab Republic because of the political instabilities in these countries as well as the job opportunities caused by large-scale development projects undertaken by Turkey. In this report, imported CL cases detected in five truck drivers transporting from Hatay to Turkmenistan, Syria, Saudi Arabia, Iran and Georgia, Uzbekistan and Azarbaijan countries were presented. The patients admitted to Mustafa Kemal University, Faculty of Medicine Dermatology Policlinic, with wound complaints on their bodies were directed to the Department of Parasitology to obtain smear samples from their wounds. The age range of the patients were 38 to 43 years. Patients with wound trail for a period ranging from one month to one year had a number of lesions varying between 2-7 and in all cases, a smear preparation was prepared from the lesions for diagnostic purposes. Clinical material obtained from five patients with pre-diagnosis of CL was firstly examined with Giemsa stain. Samples taken from the patients were inoculated into modified NNN (Novy-MacNeal-Nicolle) medium for the evaluation of the presence of the promastigotes. Promastigotes obtained from the inoculated medium were also genotyped using the ITS1 region. In all of the slides prepared from the clinical material taken from the patients amastigotes were determined. The growth of promastigotes were observed in only three of the clinical specimen inoculated media. The genotyped three species were Leishmania tropica, Leishmania infantum/donovani and Leishmania major. In this study, the importance of support for the diagnosis of different microbiological methods used in the diagnosis of leishmaniasis infection which occurred during the outbreaks of the disease has been put forward. In addition, it was aimed to draw attention to the importance of imported CL cases in our country diagnosed in five truck drivers making transportation from Hatay to Turkmenistan, Syria, Saudi Arabia, Iran, Georgia, Uzbekistan and Azerbaijan.

[Determination of imported malaria cases in Hatay by the use of molecular methods]. / Hatay'da yurt disi kaynakli sitma olgularinin moleküler yöntem kullanarak degerlendirilmesi.

Malaria, being among the most important diseases throughout history, is still an important public health problem among parasitic diseases due to increasing population movements with various reasons such as migration, war and travel. According to WHO data each year 300-350 million people get exposed to malaria, each year 1.5-2.7 million people die from malaria and also 40% of the world's population is still at risk for this disease. According to World Health Organisation (WHO) data, imported cases were not reported since 2013 in our country. However among imported cases Plasmodium falciparum malaria can be observed. The aim of this study wasto draw attention to the imported malaria cases increasing gradually and to the importance of the chemoprophylaxis in terms of malaria before travelling. In the study, male patients who have admitted to Hatay Province Malaria Center or Mustafa Kemal University Infectious Disease Department, ages between 25-60 years, were analyzed. All of the patients have worked abroad before. Patients were mostly from Sudan but there were also patients from endemic regions such as Africa, Nigeria, Pakistan, Mali island. The cases were evaluated according to age, gender and whether they had travel stories in Turkey or abroad. Blood samples taken from the patients were firstly prepared by thin and thick smear preparations and examined microscopically by staining with Giemsa stain method. Samples that were found positive by microscopic examination were impregnated on drying papers and genotyped using nested-PCR. Out of the 30 samples from patients who had traveled to endemic countries before, determined as positive by microscopical examination and genotyped by nested-PCR, 16 of them were identified as P.falciparum, six of them as P.vivax and eight of them as P.falciparum/P.vivax. The study suggested that malaria prophylaxis has to be applied before travelling to endemic countries, in return imported malaria has to be considered one of the first diseases in mind and people who will travel should be informed about this disease before travel.

Leishmaniasis in Turkey: first clinical isolation of Leishmania major from 18 autochthonous cases of cutaneous leishmaniasis in four geographical regions.

OBJECTIVE: To report isolation of Leishmania major strains obtained from 18 Turkish autochthonous cutaneous leishmaniasis (CL) patients infected with L. major between 2011 and 2014. METHODS: Initial diagnosis relied on microscopy and culture in enriched medium, prepared by adding specific amounts of liver extract, protein and lipid sources to NNN medium. Promastigotes were then transferred to RPMI medium including 10% of foetal calf serum for mass culture. Species-specific real-time PCR targeting ITS1 region of Leishmania spp. was performed using both lesion aspiration samples and cultured promastigotes. Two of 18 isolates were identified by isoenzyme analysis in the Leishmaniasis Reference Center in Montpellier, France. Each isolate was inoculated into the footpads of six mice to observe the pathogenicity of L. major. Developing lesions were observed, and the thickening of footpads was measured weekly. RESULTS: Melting curve analyses of 18 isolates showed a peak concordant with L. major, and two of them were confirmed by isoenzyme analyses as L. major zymodeme MON103. In the mouse model, acute lesions seen on day 21 were accepted as an indication of heavy infection. Severe impairments were observed on all mouse footpads over 3 weeks, which even progressed to extremity amputation. CONCLUSION: Cutaneous leishmaniasis-causing L. major was recently identified in Adana province in southern Turkey, with PCR. Our study shows that such CL cases are not limited to Adana but currently present from western to Southeastern Anatolia, and along the Mediterranean coast. The role of small mammals, the main reservoirs of L. major in Anatolia, needs to be elucidated, as do the underlying factors that cause severe clinical manifestations in L. major infections in Turkey, contrary to the infections in neighbouring countries.

Leishmaniasis recidiva cutis of the lips mimicking granulomatous cheilitis.

Leishmaniasis recidiva cutis (LRC) is an unusual form of acute cutaneous leishmaniasis. Herein, we present a case of LRC of the lips mimicking granulomatous cheilitis. An 8-year-old, Syrian child admitted with a swelling and disfigurement of his lips for 4 years. Abundant intra and extracellular Leishmania amastigotes were determined in the smear prepared from the lesion with Giemsa stain. Histopathology showed foamy histiocytes and leishmania parasites within the cytoplasm of macrophages in the epidermis and a dense dermal mixed type inflammatory cell infiltrate composed of lymphocytes, foamy histiocytes with multinucleated giant cells. On the basis of anamnestic data, the skin smears results, clinical and histopathologic findings, LRC was diagnosed. The patient was treated with meglumine antimoniate intramuscularly and fluconazole orally. Cryotherapy was applied to the residual papular lesions. The lesion improved markedly at the first month of the treatment.

Leishmaniasis in Turkey: Determination of Leishmania Species by Matrix-Assisted Laser Desorption Ionization Time-Of-Flight Mass Spectrometry (MALDI-TOF MS).

BACKGROUND: 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.

[Is the agent of cutaneous leishmaniasis in Sanliurfa changing? First cases of Leishmania major]. / Sanliurfa'da Sark Çibani Etkeni Degisiyor Mu? Ilk Leishmania Major Vakalari.

Today, almost 2 million new leishmaniasis cases are noted annually; 1.5 million of these are cutaneous (CL), and others are visceral leishmaniasis (VL). In Sanliurfa, CL cases caused by Leishmania tropica but not by other agents such as L. infantum and L. major. L. tropica is a unique parasite species in Sanliurfa and is the causative agent of anthroponotic CL (transmitted from human to vector to human). Our aim was to report 3 new CL cases due to L. major ( 2 autochthonous and 1 imported) identified in Sanliurfa. Lesion aspiration samples taken from patients were inoculated into NNN culture. Following successful isolation in NNN, promastigotes were obtained by mass culture using RPMI + 20% FCS medium. Parasites species were identified as L. major using ITS-1 PCR-RFLP analysis. This is the first report of autochthonous CL cases caused by L. major in Sanliurfa, and it is estimated that the number of such cases will increase in this region. Public health measures should be taken for L. major infections, while researchers should plan field studies to identify the vectors and reservoirs of L. major.

A real-time ITS1-PCR based method in the diagnosis and species identification of Leishmania parasite from human and dog clinical samples in Turkey.

Human visceral leishmaniasis (VL) caused by L. infantum and cutaneous leishmaniasis (CL) caused by L. tropica and L. infantum have been reported in Turkey. L. infantum is also responsible for canine leishmaniasis (CanL) and it is widely common in the country. The main aim of the present study was to design a real-time PCR method based on the internal transcribed spacer 1 (ITS1) region in the diagnosis of all clinical forms of leishmaniasis in Mediterranean, and to identify the species directly from clinical samples. Totally, 315 clinical specimens, human/canine visceral (blood, bone marrow, lymph node) and cutaneous (lesion aspiration) samples, and 51 Turkish Leishmania isolates typed by isoenzymatic method were included in the study. For optimization, DNA samples of the 34 strains were amplified by conventional ITS1-PCR and then sequenced for designing the primers and probes, allowing the species identification. Following the validation with the isolates, the test was applied on clinical samples and melting temperatures were used for genotyping. A group of PCR products were further sequenced for confirmation and assigning the inter- and intraspecies heterogeneity. The diagnosis of leishmaniasis is successfully achieved by the new real-time PCR method, and the test identified 80.43% of human and canine VL samples as L.infantum and 6.52% as L.tropica; 52.46% of CL samples as L. infantum and 26.90% as L. tropica. In 13.04% of visceral and 20.62% of cutaneous samples, two peaks were observed. However, the higher peak was found to be concordant with the sequencing results in 96.96%, in terms of species identification. The real-time ITS1 PCR assay clearly identified the leishmanial species in 81.58% of all clinical samples. Genotypic variations of Leishmania parasites in Turkey within species and intraspecies were observed, and L. tropica is also found as causative agent of human and canine VL in Turkey.