Comparison of Vitek Matrix-assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry Versus Conventional Methods in Candida Identification.

Candida species are generally identified by conventional methods such as germ tube or morphological appearance on corn meal agar, biochemical methods using API kits and molecular biological methods. Alternative to these methods, rapid and accurate identification methods of microorganisms called matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has recently been described. In this study, Candida identification results by API Candida kit, API 20C AUX kit and identifications on corn meal agar (CMA) are compared with the results obtained on Vitek-MS. All results were confirmed by sequencing internal transcribed spacer (ITS) regions of rDNA. Totally, 97 Candida strains were identified by germ tube test, CMA, API and Vitek-MS. Vitek-MS results were compatible with 74.2 % of API 20C AUX and 81.4 % of CMA results. The difference between the results of API Candida and API 20C AUX was detected. The ratio of discrepancy between Vitek-MS and API 20C AUX was 25.8 %. Candida species mostly identified as C. famata or C. tropicalis by and not compatible with API kits were identified as C. albicans by Vitek-MS. Sixteen Candida species having discrepant results with Vitek-MS, API or CMA were randomly chosen, and ITS sequence analysis was performed. The results of sequencing were compatible 56.2 % with API 20C AUX, 50 % with CMA and 93.7 % with Vitek-MS. When compared with conventional identification methods, MS results are more reliable and rapid for Candida identification. MS system may be used as routine identification method in clinical microbiology laboratories.

Genotyping and phylogenetic analysis of Giardia duodenalis isolates from Turkish children.

BACKGROUND: Giardiasis is caused by the intestinal protozoan parasite Giardia duodenalis (synonyms: G. lamblia, G. intestinalis), which is one of the most frequent parasites that infect Turkish children. However, molecular characterization of G. duodenalis in Turkey is relatively scarce. The present work aimed at genotyping G. duodenalis isolates from Turkey using molecular techniques. MATERIAL AND METHODS: In the present study, 145 fecal samples from children were collected to search for the presence of Giardia by microscopy and PCR screening. PCR generated a 384 bp fragment for ß-giardin. The PCR products were sequenced and the sequences were subjected to phylogenetic analysis by using PHYLIP. RESULTS: Based on the phylogenetic analysis of the sequences, assemblage A, B, and mixed subtypes were determined. Of 22 isolates, 11 were identified as assemblage A (50%), 7 were assemblage B (31.8%), and 4 were assemblage AB (18.2%). Association between G. duodenalis assemblages and the epidemiological data was analyzed. No correlation was found between symptoms and infection with specific assemblages (P>0.05), but we found statistically significant association between age and the assemblage AB (P=0.001). CONCLUSIONS: The association between G. duodenalis and the epidemiologic data were analyzed. Since assemblage A is the more prevalent subgroup compared with assemblage B, this subgroup might be responsible for common Giardia infections in Turkey. This is the first study that included a detailed phylogenetic analysis of Giardia strains from Turkey.

Evaluation of immunochromatographic test for the detection of antibodies against Echinococcosis granulosus.

BACKGROUND: Echinococcosis in humans is a disease caused by the larvae of Echinococcus granulosus (E. granulosus) and Echinococcus multilocularis (E. multilocularis). Serological tests are valuable, especially in the clarification of unexplained clinical findings and imaging methods. For this reason, indirect hemagglutination (IHA), latex agglutination, immunoelectrophoresis, immunoblotting, immuno-enzymatic tests, indirect fluorescence antibody test (IFAT), and enzyme-linked immunosorbent assay (ELISA) are used. The purpose of this study was to investigate the value of an immunochromatographic test (ICT) specific for E. granulosus antibodies in the diagnosis of echinococcosis. MATERIAL/METHODS: ICT evaluated 102 cases of cystic echinococcosis, 38 cases of other parasitic diseases, and 50 healthy individuals. ELISA (DRG, Germany) that detects IgG antibodies specific for E. granulosus was used as the reference method. RESULTS: The sensitivity, specificity, and positive and negative predictive values of ICT were 96.8%, 87.5%, 98.9%, and 70%, respectively. Diagnostic value was 96.1%. No significant differences and high degrees of agreement were found between ELISA and immunochromatographic test for cystic echinococcosis. Serum samples included 4 taeniasis, 2 leishmaniasis, and 2 healthy individuals were diagnosed to be positive with immunochromatographic test. CONCLUSIONS: The ability of test to give fast results without need for equipment, devices, and specific storage conditions is an advantage. This test may be used due to its advantages in endemic regions for screening and diagnostic purposes.

Long pentraxin-3 measured at late phase associated with GRACE risk scores in patients with non-ST elevation acute coronary syndrome and coronary stenting.

OBJECTIVES: We analyzed pentraxin 3 (PTX3) levels and the relation of PTX3 levels with GRACE risk scores in 39 patients with non-ST elevation acute coronary syndrome (ACS) and stabile angina after stenting. STUDY DESIGN: Seventeen patients with ACS and 22 patients with stabile angina who underwent coronary stenting were included in the study. PTX3 levels were measured serially at admission, at the 8th hour and at the 24th hour after stenting. RESULTS: While diabetes and hypertension were more frequent in the stabile angina group, leukocyte counts were significantly higher in the ACS group. PTX3 levels measured at the 8th hour were significantly higher in the ACS group compared to the stabile angina group (p=0.003). Strong correlations were observed between 24th hour PTX3 levels and GRACE scores calculated for risk of death and death/MI at admission (in-hospital/to 6 months), and for risk of death/MI at discharge to 6 months (R=0.571, p=0.01, R=0.564, p=0.01; R=0.558, p=0.02, R=0.512, p=0.03; R=0.653, p=0.004, respectively). CONCLUSION: The serum PTX3 levels may provide important information for the early risk stratification of patients with ACS who underwent coronary stenting.

Seroprevalence of Toxoplasma gondii, rubella and cytomegalovirus among pregnant women in western region of Turkey.

PURPOSE: Prenatal screening for antibodies to Toxoplasma gondii (T. gondii), rubella virus and cytomegalovirus (CMV) infectious agents is an important tool in this process. The aim of this study was to determine the seroprevalence of these infections through antenatal screening in Kocaeli region. MATERIAL AND METHODS: 1972 samples of sera were tested for antibodies to TORCH agents known to cause serious congenital infections: T. gondii, rubella, CMV. Anti-Toxoplasma, anti-rubella and anti-CMV IgM and IgG antibodies were assayed by ELISA method using Abbott kits (Axsym, Abbott, USA) according to the manufacturer's instructions. RESULTS: Of 1972 pregnant women, seropositivity for anti-toxoplasma IgG antibody was found in 952 (48.3%), while 8 (0.4%) of the subjects tested were positive for the anti-Toxoplasma IgM antibody only, and 31 (1.6%) of the subjects tested were positive for anti-Toxoplasma IgG+IgM antibodies together. The seropositivities for anti-rubella IgG, IgM and IgG+IgM together were found in 1896 (96.1%), 4 (0.2%) and 35 (1.8%) of the pregnant women, respectively. The seropositivities for anti-CMV IgG, IgM and IgG+IgM together were found in 1900 (96.4%), 13 (0.7%) and 37 (1.9%) of the pregnant women, respectively. CONCLUSION: Widespread population screening may contribute to the prevention of congenital infections due to TORCH agents. Because of the high seropositivity of T. gondii, rubella and CMV in pregnant women, preventive measures should be taken.

Primary hydatid disease in adductor muscles.

Primary muscular Echinococcus infection is very rare without involvement of thoracic and abdominal organs. In this case a 31-year-old man who had a growing mass in the postero-medial part of his right thigh was examined. The mass was diagnosed as hydatid cyst using ultrasound, magnetic resonance imaging (MRI) and serological tests. It was removed surgically and there has been no recurrence one year after the surgery. The MRI imaging characteristics may differ depending on the life cycle stage of the parasite. In this case report, we discuss the imaging characteristics of the muscular hydatid cyst with special emphasis on the MRI findings. In regions where hydatidosis is endemic, a mass found in body muscles should be considered as a muscular hydatid cyst.

The schizophrenia and Toxoplasma gondii connection: infectious, immune or both?

INTRODUCTION: Recent research has suggested a possible link between toxoplasmic agents and schizophrenia. We aimed to assess this by measuring Toxoplasma gondii-associated antibodies in schizophrenia patients and controls METHODS: We used a commercially available enzyme-linked immunosorbent assay (ELISA) kit to measure the level of immunoglobulin G (IgG) and IgM antibodies in serum samples from schizophrenia patients (n=40) and from a group of non-schizophrenic control subjects (n=37) RESULTS: Among schizophrenic patients, 16 (40%) showed IgG seropositivity and two (5%) showed IgM seropositivity. Among the control group, five (13.5%) were found have IgG seropositivity and one (2.7%) showed IgM seropositivity. In our study we found that IgG T gondii antibodies were significantly higher in schizophrenia patients compared with controls CONCLUSIONS: This study supports the theory that toxoplasmic agents may have a role in the aetiology of schizophrenia.

[Evaluation of Malaria Cases that Were Detected in Kocaeli Province During 2008 Through 2013]. / Kocaeli Ilinde 2008-2013 Yillari Arasinda Saptanan Sitma Olgularinin Degerlendirilmesi.

OBJECTIVE: Malaria is still a serious public health problem around the world and in our country. In this study, we examined the epidemiology of malaria cases retrorespectively by using the surveillance data provided by the Malaria Control Unit of the Infectious Disease Division of Kocaeli Health Directory, from the years of 2008 to 2013. Our aim was to compare our findings with the findings of the past studies and provide support to the future surveillance studies. METHODS: A total of 10008 periferic blood samples were examined in this surveillance based study. The cases were evaluated according to age groups, gender, month during which the infection was detected and the origin of district. RESULTS: The species of malaria parasites were detected in 27 samples of which 51.9 % (n=14) was Plasmodium vivax and 48.1% (n=13) was Plasmodium falciparum. Among 27 cases 77.8% (n=21) of the cases were male and 22.2% (n=6) were female. 96.3% of the cases were above the age of 15. All cases of Plasmodium vivax were consisted of the workers who came to Kocaeli from Southeastern Anatolia Region. CONCLUSION: Malaria cases detected in our region have displayed a relatively meaningful decrease in comparison to past. While Plasmodium vivax was dominating infecting agent in the past, in recent years Plasmodium falciparum appeared to be infecting nearly half of the cases. These cases were imported cases coming to Kocaeli from other countries. It is important to take the necessary precautions for timely diagnosis of imported cases and to prevent its spread in the area.

Interaction between caspofungin or voriconazole and cefoperazone-sulbactam or piperacillin-tazobactam by in vitro and in vivo methods.

Immunosuppressive patients are at risk of fungal and bacterial infections. Therefore, these patients receive prophylactic, preemptive, empirical or target antifungal and concomitant antibiotic therapy. To this end, caspofungin (CAS) or voriconazole (VRC) antifungals and cefoperazone-sulbactam (CPZ/SAM) or piperacillin-tazobactam (PIP/TAZ) antibiotics may be used. Here, we aimed to investigate the interaction between these antifungals and antibiotics by in vitro and in vivo methods. The interaction was tested by chequerboard analysis and fractional inhibitory concentration index (FICI). It was also tested in a neutropenic mice-invasive candidiasis model and evaluated by fungal burden in kidney tissue of infected animals from the first day to the fifth day of treatment with 24 h intervals. A synergism was detected between CAS and CPZ/SAM (FICI = 0.1) and PIP/TAZ (FICI = 0.3). Fungal burden in tissues of drug-treated mice was reduced compared with controls in a time-dependent manner. In comparison with CAS-alone treated group, there were 1.32 log10 reductions of fungal burden in CAS + CPZ/SAM (p = 0.002) and in CAS + PIP/TAZ group (p = 0.14). The same interactions were not found with VRC and antibiotics. CPZ/SAM had stronger synergistic interaction with CAS than PIP/TAZ. The mechanism of synergism is not well understood. This is most likely due to an increase in the anticandidal effect of CAS plus antibiotics.

Plasma antiphospholipid antibody levels in age-related macular degeneration.

PURPOSE: To investigate the association of age-related macular degeneration (AMD) with plasma antiphospholipid antibody levels. METHODS: This prospective study included 19 patients diagnosed as having dry-type AMD, 23 patients with exudative-type AMD, and 25 control subjects. Venous blood samples of the participants were obtained. Anticardiolipin antibodies (aCL) isotypes IgG and IgM were measured by means of an enzyme-linked immunosorbent assay. Lupus anticoagulant (LA) antibodies were measured by the dilute Russell viper venom time screen test. RESULTS: The mean aCL IgG concentration in patients with exudative-type AMD was significantly higher than in patients with dry-type AMD and control subjects. The mean ± SE of aCL IgG levels in patients with exudative-type AMD and dry-type AMD and control subjects was 5.46 ± 1.26; 2.55 ± 0.78; and 0.32 ± 0.1, respectively. The mean aCL IgM levels and LA levels in the 3 groups were not statistically different. CONCLUSIONS: Our findings suggest that elevated levels of serum aCL, a risk factor for cardiovascular and cerebrovascular diseases, may be associated with exudative-type AMD.

Macrolide and tetracycline resistance and emm type distribution of Streptococcus pyogenes isolates recovered from Turkish patients.

The aims of this study were to determine the susceptibilities to macrolide and tetracycline antibiotics and emm type distribution of Streptococcus pyogenes strains isolated in the Kocaeli University Hospital, Turkey. A total of 127 S. pyogenes clinical isolates were tested. Eleven (9%) isolates were resistant to erythromycin, and 23 (18%) isolates were resistant to tetracycline. Ten of the erythromycin-resistant isolates were also resistant to tetracycline. By the triple-disk test, all erythromycin-resistant isolates showed the inducible macrolide-lincosamide-streptogramin-C phenotype and harbored erm(TR) gene. tet(O) was the most common tetracycline resistance gene. Among erythromycin-tetracycline coresistant isolates, seven harbored the tet(O) gene. emm 4, emm 1, emm 2,114, and emm 89 were the most common emm types. These isolates were more susceptible to erythromycin. There was considerable emm type heterogeneity in macrolide or tetracycline resistant isolates. According to our knowledge, this is the first study in which emm type distribution is investigated in Turkey. More comprehensive studies are needed to obtain true information about the epidemiology of macrolide and tetracycline resistance and emm type distribution in Turkey.

Routine using pattern and performance of diagnostic tests for tuberculosis on a university hospital.

INTRODUCTION: Nucleic acid amplification tests to detect Mycobacterium tuberculosis in clinical specimens are used increasingly as a laboratory tool. We aimed to investigate the routine using pattern and the effects on therapeutic decision of diagnostic tests for tuberculosis in our hospital. METHODS: In this descriptive study, we investigated retrospectively the routine using pattern and the effects on therapeutic decision of diagnostic tests for tuberculosis. Patients with discordant results were clinically evaluated retrospectively by a chest physician. Samples were tested for the presence of M. tuberculosis by a smear technique, M. tuberculosis culture growth technique (Löwenstein-Jensen and/or BACTEC-960), and IS6110 polymerase chain reaction (PCR). RESULTS: Culture positivity was 7.2% (83 of 1159 patients). In total, 198 (62.4%) were tested with PCR, acid-fast bacilli, and culture. On the basis of culture results as a gold standard, sensitivity, specificity, positive predictive value, and negative predictive value of PCR were 46%, 89%, 23%, and 93.5%, respectively. CONCLUSIONS: Selection of appropriate patients for further testing and exclusion of low-risk patients from microbiologic testing by experienced clinicians may help to optimize the positive predictive value of PCR.

[Case report: strongyloidiosis with chronic abdominal pain]. / Olgu Sunumu: Kronik Karin Agrisiyla Seyreden Strongyloidosis.

The case was presented here in order to point out that an immunocompetent child might have Strongyloidiosis infection that might be misdiagnosed. A 9 year old male patient who had chronic abdominal pain with a feeling of weakness was treated several times for urinary tract infection. He had never been tested for the presence of parasites. After the patient's complaints occurred again, he presented at our hospital. Strongyloides stercoralis larvae were observed in his feces by microscopy. Albendazol (400 mg/day for three days) was prescribed. After 10 days, the feces of the patient was reexamined and no Strongyloides stercoralis larvae were detected. For this reason, it is important to investigate the possibility of intestinal parasitic infections in children with chronic abdominal pain.

[The investigation of the presence of antibodies for Cryptosporidium spp. in fecal samples using ELISA]. / Diskida Cryptosporidium spp. Antijenlerinin ELISA ile Arastirilmasi.

In this study, difficulties in the effective diagnosis of cryptosporidiosis were considered. Fecal samples of 80 patients who applied to our clinic with gastrointestinal complaints were analyzed for the presence of cryptosporidiosis by the acid-fast staining method and ELISA kit (R-Biopharm, Germany) which detects the presence of Cryptosporidium spp. specific antibodies. The results of ELISA were compared with the results of the acid-fast stain and the question of whether ELISA can be used for effective diagnosis of cryptosporidiosis was investigated. Out of 80 fecal samples, 3 samples (3.75 %) were found to be positive for oocysts of Cryptosporidium spp. with the acid-fast stain and 5 samples (6.25 %) were found to be positive with ELISA. Three samples were found to be positive with both methods. In this study, the sensitivity, specificity, negative preductivity and positive preductivity with Cryptosporidium ELISA kit were 60%, 100%, 97.4% and 100%, respectively. These findings showed us that. ELISA should be considered as the choice of method for the diagnosis of cryptosporidiosis in patients who have had diarrhea for a long-period of time, even though the method is more costly.

[Distribution of intestinal parasites among patients who presented at the parasitology laboratory of the Kocaeli University School of Medicine Hospital]. / Kocaeli Universitesi Tip Fakültesi Hastanesi Parazitoloji Laboratuvarina Basvuran Hastalarda Bagirsak Parazitlerinin Dagilimi.

In this study, 5178 fecal samples collected from patients who applied to our university during May 2003 through June 2005 were examined for the presence of intestinal parasites. Out of 5178 samples, 4560 of them were collected with cellophane tape preparations. Fecal samples were examined using native- Lugol and the trichrome staining method after sedimentation by formal-ethyl acetate concentration. The prevalence of intestinal parasites was evaluated with respect to age, gender and the months in which cases were seen. One or more parasites were found in 553 (10.67%) of the 5178 stool samples. Of these, 39.8% were female and 60.2, male. The distribution of the intestinal parasites was as follows; G. intestinalis in 138 cases (24.9%), E. vermicularis in 129 cases (23.3%), D. fragilis in 10 cases (1.8%), I. bütschlii cysts in 26 cases (4.7%), Taenia spp in 17 cases (3.07%) and Chilomastix mesnili in one case (0.18%). The rate of the presence of intestinal parasites in the 0-14 age group was higher than that of other groups. The incidence of parasitic diseases was lower in this study as compared to the results previously reported from our hospital. In our region intestinal parasites were predominantly protozoan whereas a lower rate of helminth isolations was noticeable. Parasitic infections continue to be an important public health problem in our country.

[The frequency of the presence of intestinal parasites in students of Arslanbey Primary School]. / Arslanbey Ilkögretim Okulu Ogrencilerinde Bagirsak Parazitlerinin Görülme Sikligi.

We investigated the frequency of the presence of intestinal parasites and the factors affecting this frequency in an elementary school. We tried to discover whether there was a relationship between the presence of intestinal parasites and sanitary habits such as hand washing, toilet flushing, and use of human manure, cleanness of drinking water and also the educational level of the parents. A total of 114 students participated in this study. Samples were collected using cellophane tape preparations. All fecal samples were examined using native-Lugol and the trichrome staining. Fecal samples taken from 111 students were examined, 37 (33.3%) of them contained one or more parasites. The parasites encountered most often were E. vermicularis 16 (14.4%), G. intestinalis 10 (9.0%), and E. coli 4 (3.6%). In addition children carrying intestinal parasites complained about abdominal pain (22 students), itchy nose and the accumulation of saliva around their mouths (9 students). Although no meaningful relationship was found between the use of human manure/toilet flushing with the presence of parasites, the educational level of the parents, cleanness of the drinking water and hand washing habits seem to be related to the presence of parasites. Our findings indicate that elementary students should be educated in the prevention of parasitic infections.

[The prevalence of cryptosporidiosis in children who were diagnosed with leukemia and lymphoma]. / Lösemi ve Lenfoma Tanisi Alan Cocuklarda Cryptosporidiosis Prevalansi.

Cryptosporidium spp. is known to cause heavy diarrhea especially in immunosuppressed patients. In this study, eighty nine leukemia and lymphoma patients between the ages of 1 to 14 were studied for the prevalence of Cryptosporidiosis using both ELISA (Cryptosporidium Rida Screen, R-Biopharm, Germany) and the Kinyoun acid-fast staining method. These patients were sent to us by the Hematology-Oncology department where they were diagnosed with leukemia and lymphoma. Cryptosporidium spp. were detected in 11 patients (12.35%) with ELISA and in 7 patients (7.86%) with the Kinyoun acid fast stain. No cryptosporidiosis was detected in the control groups of 60 patients with neoplasia but without diarrhea. The distribution of Cryptosporidium among positive samples were 7 (14.8%) in patients who were diagnosed with ALL, 3 (10%) in patients who were diagnosed with KML, and 1 (8.3%) in patients who were diagnosed with solid tumors. Sixty-five patients (73.03%) had a fever, 43 patients (48.31%) were vomiting and 58 patients (65.16%) had stomach pain. Except for two, all the patients responded positively to paromomycin treatment. Those two patients responded positively to azitromycine treatment. We suggest that when considering cryptosporidiosis in children with cancer, the use of a more sensitive and specific method such as ELISA- in addition to the acid fast stain should be considered.

[Seroprevalence of visceral leishmaniasis in stray dogs in Kocaeli]. / Kocaeli Sokak Köpeklerinde Visseral Leishmaniasis Seroprevalansi.

Dogs are the most important reservoir of visceral leishmaniasis (VL). A male child who lives in Köseköy in Kocaeli was diagnosed with VL. Since this child had never been outside Kocaeli, serum samples of 65 stray dogs were analyzed for canine VL using the indirect fluorescence antibody test (IFAT) and ELISA. Two dogs (3.07%) tested positive with both ELISA and IFAT. Leishmania amastigotes were observed in the lymph aspiration material from one of them. Growth was observed in NNN medium inoculated with lymph aspiration material from the other dog. This was the first study investigating canine VL prevalence in our city and plans were made for control of the disease.

[Cystic echinococcosis in Turkey from 2001-2005]. / Türkiye'de 2001-2005 Yillari Arasinda Kistik Ekinokokkozis.

Cystic echinococcosis (CE) caused by the metacestode form of Echinococcus granulosus is a major public health problem especially in animal-raising regions of the world. In the present study, CE cases were determined during 2001-2005 by investigating different hospital and health directorship documents and Health Ministry documents, retrospectively. Our results show that there were 2534 (13.13%) cases in the Marmara region; 2114 (16.94%), in the Aegean region; 2578 (16.09%), Mediterranean region; 5404 (38.57%), in the Middle Anatolian region; 428 (5.70%), in the Black Sea region; 844 (6.80%), in the eastern Anatolian region; and 887 (2.75%), in the southeastern Anatolian region making a total of 14,789 CE cases. Finally, it has been determined that the patients were hospitalized for a total of 149,464 days.