HEV seroprevalence in blood donors in Turkey by two commercial total anti-HEV Ab ELISA kits.

Previous hepatitis E virus (HEV) seroprevalence studies in Turkey have shown high variabilities, leading to conflicting results. We aimed to re-evaluate HEV seroprevalence among blood donors in Turkey using the Wantai (Beijing, China) and the Dia.Pro (Milan, Italy) total anti-HEV antibody (Ab) enzyme-linked immunosorbent assay (ELISA) kits and compare their performances and to investigate the presence of HEV RNA in blood donors. Serum total anti-HEV antibodies were determined in a total of 2011 volunteer blood donor samples collected from different regions of Turkey (807 from Ankara, 243 from Kayseri, 284 from Izmir, 200 from Malatya, 200 from Kahramanmaras, and 277 from Van). HEV RNA was evaluated by a real-time polymerase chain reaction in a total of 272 anti-HEV seropositive samples. The country-wide HEV seroprevalence was calculated as 11.5% (Dia.Pro) and 12.2% (Wantai) with seropositivity rates of 12.0%-12.5% in Ankara, 7.4%-8.2% in Kayseri, 14.5%-15.5% in Malatya, 8.1%-8.8% in Izmir, 15.0%-16.0% in Kahramanmaras, and 12.6%-13.4% in Van by Dia.Pro and Wantai kits, respectively. The lowest detectable Ab concentrations were 0.16 and 0.14 units/mL WHO, for the Dia.Pro and the Wantai assays, respectively, showing no significant difference between assays. HEV RNA was not detected in any of the anti-HEV seropositive samples. Compared with previous studies, HEV was shown to have a higher overall seroprevalence in Turkey. Despite its limitation, the current study represents the most comprehensive HEV seroprevalence study in Turkey performed with two different commercial ELISA assays with high sensitivities so far. Further investigation is required to determine HEV genotypes in Turkey.

Effectiveness of quercetin in an experimental rat model of allergic rhinitis.

We aimed to investigate whether quercetin had a therapeutic effect in an experimental rat model of allergic rhinitis. The study was conducted with 35 rats, which were randomly assigned into 4 groups: group 1 (n = 5), sham group; group 2 (quercetin group, n = 10) received 80 mg/kg day quercetin; group 3 (steroid group, n = 10) received steroid (mometasone furoate); and group 4 (control group, n = 10), received ovalbumin alone. Rats were sensitized by administration of ovalbumin on alternate days over 14 days via an intraperitoneal route. On day 15, in addition to ovalbumin via an intranasal route, quercetin and steroid were given over 7 days to the corresponding groups. All rats were then sacrificed and nasal turbinates were evaluated histopathologically, and serum total IgE and ovalbumin (OVA)-specific IgE values were measured before and after treatment. A significant increase in OVA-specific IgE values was detected in all groups except sham group. A significant increase was detected in post-treatment total IgE levels in the control group, while no significant change was detected in the sham, quercetin, and intranasal steroid groups. On histopathological evaluation, it was observed that findings of allergic rhinitis were suppressed in the quercetin group when compared to the control group. In immunohistochemical evaluation, it was detected that COX-2 and VIP expressions were weaker in the quercetin group compared to the control group. Based on these findings, we conclude that quercetin was effective in allergic rhinitis induced by ovalbumin in rats both histopathologically and serologically.

Is surveillance for colonization of carbapenem-resistant gram-negative bacteria important in adult bone marrow transplantation units?

BACKGROUND: The aim of this study was to investigate the rate of carbapenem-resistant gram-negative bacilli (CRGNB) colonization and to analyze the risk factors associated with CRGNB colonization. METHODS: This prospective study was conducted in adult patients hospitalized in hematopoietic stem cell transplantation (HSCT) units over a period of 8 months. Rectal swab samples were obtained from each participant every Monday, and patients CRGNB positive on admission were excluded. RESULTS: Of 185 participants, the median age was 47 years, and 59.5% were men. CRGNB colonization was detected in 21 (11.4%) patients. The most commonly isolated CRGNB were Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa. Multivariate analysis revealed that busulfan use (11.9 times), fludarabine use (6.4 times), transfer from another hospital (7.8 times), transfer between units (9.3 times), and central venous catheterization (5.1 times) were risk factors for CRGNB colonization. During the study period, febrile neutropenia (FN) developed in 9 (56.2%) of the 21 colonized patients, and 1 patient died. CONCLUSIONS: Screening of patients for CRGNB colonization may have a role in preventing the spread of CRGNB. However, the empirical antimicrobial treatment for FN in patients with CRGNB colonization did not change, and their mortality rates were similar.

Antifungal Activity of Propolis Against Yeasts Isolated From Blood Culture: In Vitro Evaluation.

BACKGROUND: Due to the failure of available antifungal agents in the treatment of candidemia and the toxic activities of these drugs, a lot of researches are being conducted to develop new nontoxic and effective antifungal agents for optimal control of fungal pathogens. The aim of this study is to evaluate the in vitro antifungal activity of propolis against yeasts isolated from the blood cultures of intensive care unit patients. METHODS: Seventy-six strains were included in this study. The in vitro antifungal activity of propolis, fluconazole (FLU), and itraconazole (ITR) was investigated by the microdilution broth methods (CLSI guidelines M27-A3 for yeast). The propolis sample was collected from Kayseri, Turkey. RESULTS: Of the 76 isolates, 33 were identified as Candida albicans while 37 were C. parapsilosis, three were C. tropicalis, and three were identified as C. glabrata. The geometric mean range for MIC (µg/ml) with regard to all isolates was 0.077 to 3 µg/ml for FLU and ITR, and 0.375 to 0.70 µg/ml for propolis. It was shown that propolis had significant antifungal activity against all Candida strains and the MIC range of propolis was determined as 0185 to 3 µg/ml. CONCLUSION: This study demonstrated that propolis had significant antifungal activity against yeasts isolated from blood culture compared with FLU and ITR. The propolis MIC in azole-resistant strains such as C. glabrata was found lower than the FLU MIC.

Evaluation of the Abbott Real Time HCV genotype II assay for Hepatitis C virus genotyping.

OBJECTIVE: The determination of HCV genotypes and subtypes is very important for the selection of antiviral therapy and epidemiological studies. The aim of this study was to evaluate the performance of Abbott Real Time HCV Genotype II assay in HCV genotyping of HCV infected patients in Kayseri, Turkey. METHODS: One hundred patients with chronic hepatitis C admitted to our hospital were evaluated between June 2012 and December 2012, HCV RNA levels were determined by the COBAS® AmpliPrep/COBAS® TaqMan® 48 HCV test. HCV genotyping was investigated by the Abbott Real Time HCV Genotype II assay. With the exception of genotype 1, subtypes of HCV genotypes could not be determined by Abbott assay. Sequencing analysis was used as the reference method. RESULTS: Genotypes 1, 2, 3 and 4 were observed in 70, 4, 2 and 24 of the 100 patients, respectively, by two methods. The concordance between the two systems to determine HCV major genotypes was 100%. Of 70 patients with genotype 1, 66 showed infection with subtype 1b and 4 with subtype 1a by Abbott Real Time HCV Genotype II assay. Using sequence analysis, 61 showed infection with subtype 1b and 9 with subtype 1a. In determining of HCV genotype 1 subtypes, the difference between the two methods was not statistically significant (P>0.05). HCV genotype 4 and 3 samples were found to be subtype 4d and 3a, respectively, by sequence analysis. There were four patients with genotype 2. Sequence analysis revealed that two of these patients had type 2a and the other two had type 2b. CONCLUSION: The Abbott Real Time HCV Genotype II assay yielded results consistent with sequence analysis. However, further optimization of the Abbott Real Time HCV Genotype II assay for subtype identification of HCV is required.

Evaluation of chromogenic agar, [corrected] VITEK2 YST and VITEK® MS for identification of Candida strains isolated from blood cultures.

The aim of this study is to compare conventional methods, chromogenic agar, [corrected] VITEK2 YST card and VITEK®MS system for the identification of Candida strains isolated from blood cultures. Fifty-four strains were identified according to conventional methods, chromogenic agar, [corrected] VITEK2 YST card and VITEK®MS. Sequencing was used as the reference method. The 54 strains included 32 Candida parapsilosis, 19 Candida albicans, 1 Candida glabrata and 2 Candida tropicalis according to the reference method. One C. albicans and one C. glabrata isolate were misidentified as C. parapsilosis by chromogenic agar. [corrected]. Two C. parapsilosis and three C. albicans isolates were misidentified by VITEK2 YST card. Chromogenic agar, [corrected] VITEK2 YST card and VITEK®MS identified correctly 96.2%, 90.7% and 100% of all strains, respectively. We found that the chromogenic agar, [corrected] VITEK2 YST card and VITEK®MS system are easy, rapid and accurate alternative methods for the identification of yeast species in the clinical microbiology laboratory.

Investigation of the relationship between virulence factors and genotype of Candida spp. isolated from blood cultures.

INTRODUCTION: The aim of study was to investigate the virulence factors of phospholipase, proteinase, esterase production and biofilm formation in Candida species isolated from patients with candidemia, and to assess their relationship with Candida genotypes derived after repetitive sequence-based polymerase chain reaction (rep-PCR) fingerprinting. METHODOLOGY: Fifty-two strains were identified to species level according to conventional methods and sequencing. The DiversiLab system was used for the genotyping. Enzyme activities and biofilm formation were evaluated using microbiological methods. RESULTS: The 52 strains were identified as follows: 29 C. parapsilosis, 19 C. albicans, 2 C. glabrata, and 2 C. tropicalis. Phospholipase and proteinase activities were observed to have statistically significant differences between C. albicans and non-albicans Candida (NAC) strains (p < 0.05), with C. albicans strains showing higher virulence. Rep-PCR revealed eight major genotypes (A-H).The 19 C. albicans and the 33 non-albicans Candida isolates yielded seven (A-G) and four (A, B, C, H) genotypes, respectively. C. albicans strains were not shown to have a predominant genotype and showed higher phospholipase and proteinase activitiy than did NAC, regardless of genotype. Genotype H (52%) was the predominant genotype for the NAC including 27 C. parapsilosis strains, but the majority of strains showed low virulence. CONCLUSIONS: NAC species were the most common causative agent for candidemia. Genotyping showed low transmission of C. albicans strains, but transmission of C. parapsilosis was high. In candidemia, several Candida virulence factors may be responsible at the same time. However, different genotypes of Candida strains showed different virulence activity.

Is rapid antibacterial susceptibility testing medium reliable for routine laboratory practices?

OBJECTIVE: Early detection of antibiotic susceptibility profile of the isolates has critical importance in terms of immediate beginning of the appropriate treatment and increasing of treatment success, such as meningitis, bacteriemia and sepsis. In the present study, it was aimed to compare the antibiotic susceptibility results of Quicolor (Salubris Inc., Massachusetts, USA) and standard disk diffusion method. METHODS: One hundred twenty three isolates were included in this study (80 Enterobacteriaceae, 15 Staphylococci and 28 nonfermentative Gram-negative bacteria). Antibiotic susceptibility in clinical isolates was evaluated using Mueller-Hinton (MH) agar and Quicolor (ES and NF) agar plates. RESULTS: For Enterobacteriaceae, frequency of total concordance, major error, and minor error between the tests were found as 96.8%, 0.8%, and 2.4%, respectively. For Staphylococci, frequency of total concordance, major error, and minor error among the tests were found as 95.7%, 3.5%, and 0.8%, respectively. For non fermentative bacteria, frequency of total concordance, major error, and minor error among the tests were found as 83.9%, 9.6%, and 6.4%, respectively. CONCLUSIONS: Quicolor media provided reliable susceptibility results in enteric bacteria and Staphylococci. However, further studies including higher number of nonfermentative bacteria are required to determine whether the chromogenic media are appropriate for this group of bacteria.

Interdigital foot infections: Corynebacterium minutissimum and agents of superficial mycoses.

Interdigital foot infections are mostly caused initially by dermatophytes, yeasts and less frequently by bacteria. Erythrasma caused by Corynebacterium minutissimum can be confused with superficial mycoses. The aim of the study was to determine the prevalence of the etiologic agents of superficial mycoses and the frequency of Corynebacterium minutissimum in interdigital foot infections. All the samples obtained from the 121 patients with interdigital foot infections were examined directly with the use of 20% potassium hydroxide mounts and Gram stain under the microscope and cultured on Sabouraud's dextrose agar plates. In identification of superficial mycoses, the rate was found to be 14% with the cultural method and 14% with direct microscopic examination. Using a combination of direct microscopic examination and culture, a 33.8% ratio was achieved. In the culture of these samples, the most isolated factor was Trichophyton rubrum (33.7%). In 24 of the patients (19.8%) Corynebacterium minutissimum was detected by Gram staining, in 6 of these patients Trichophyton rubrum was found, Trichophyton mentagrophytes was found in 2 and Trichosporon spp. was found in 1. The examination of interdigital foot lesions in the laboratory, the coexistence of erythrasma with dermatophytes and yeast should be considered.

Interdigital foot infections: Corynebacterium minutissimum and agents of superficial mycoses

Interdigital foot infections are mostly caused initially by dermatophytes, yeasts and less frequently by bacteria. Erythrasma caused by Corynebacterium minutissimum can be confused with superficial mycoses. The aim of the study was to determine the prevalence of the etiologic agents of superficial mycoses and the frequency of Corynebacterium minutissimum in interdigital foot infections. All the samples obtained from the 121 patients with interdigital foot infections were examined directly with the use of 20% potassium hydroxide mounts and Gram stain under the microscope and cultured on Sabouraud's dextrose agar plates. In identification of superficial mycoses, the rate was found to be 14% with the cultural method and 14% with direct microscopic examination. Using a combination of direct microscopic examination and culture, a 33.8% ratio was achieved. In the culture of these samples, the most isolated factor was Trichophyton rubrum (33.7%). In 24 of the patients (19.8%) Corynebacterium minutissimum was detected by Gram staining, in 6 of these patients Trichophyton rubrum was found, Trichophyton mentagrophytes was found in 2 and Trichosporon spp. was found in 1. The examination of interdigital foot lesions in the laboratory, the coexistence of erythrasma with dermatophytes and yeast should be considered.

Francisella tularensis bacteremia: report of two cases and review of the literature.

Isolation of Francisella tularensis in blood culture is exceedingly rare and has been reported infrequently in Europe; a literature review showed 28 documented cases. Herein we report the first cases of bacteremic F.tularensis pneumonia in immunocompetent individuals in Turkey.

Moxifloxacin-associated neutropenia.

A 32-y-old woman presented with pneumonia. Treatment was started with moxifloxacin. On day 2 of moxifloxacin treatment the patient developed neutropenia. After discontinuing the moxifloxacin, neutrophil counts were normal on day 4. Clinicians should be aware of the possibility of this adverse effect in patients treated with moxifloxacin.

Molecular characterization of hepatitis A virus isolated from acute infections in Turkey.

BACKGROUND/AIMS: Hepatitis A virus is a global public health problem, especially in developing countries, and the most common cause of hepatitis in childhood. Hepatitis A virus is a single- stranded positive RNA virus subdivided to 6 genotypes (3 human,3 simian). The aim of this study was to determine the prevalent genotype in Turkey using sera of acute hepatitis A virus-infected patients from different geographical regions of the country. MATERIALS AND METHODS: Sera of 137 patients with acute hepatitis A virus from different geographical regions were collected for phylogenetic analysis. The VP1-2A region of the hepatitis A virus genome was amplified by real-time-polymerase chain reaction in 76 patients where possible. Amplified polymerase chain reaction fragments were sequenced, and phylogenetic analysis was done together with other reference hepatitis A virus sequences obtained from GenBank database. RESULTS: Sequencing and phylogenetic analysis of the VP1-2A junction of hepatitis A virus showed that the most prevalent genotype in Turkey is IB (100%). Comparison of Turkish isolates and reference sequences of genotype IB showed a similarity of 94.9%. The same comparison was done between Turkish isolates and reference hepatitis A virus genotype IB and HM175, and it was found that similarity between them ranged from 93.0-95.9%. When Turkish isolates were compared according to Mean Percentage Nucleotide Distance analysis, similarity ranged between 95.3%-100%. CONCLUSIONS: Phylogenetic analysis pointed out that all Turkish isolates belong to genotype IB. Sequence analysis is a useful tool in revealing hepatitis A outbreaks, and allows us to detect and distinguish the presence of epidemic and small outbreaks.

[DNA extraction and identification of Trichophyton rubrum by real-time polymerase chain reaction from direct nail scraping specimens of patients with onycomycosis]. / Onikomikozlu Olgularin Direkt Tirnak Kazinti Örneklerinden DNA Izolasyonu ve Gerçek Zamanli Polimeraz Zincir Reaksiyonu ile Trichophyton rubrum Tanimlanmasi

Trichophyton rubrum is the most frequently encountered dermatophyte species causing onichomycosis. The routine diagnosis of dermatophytes depends on the direct microscopic examination (DME) and culture methods, however due to the phenotypic identification problems related to those agents, the molecular methods come into question. The aim of this study was to evaluate the diagnostic performance of real-time polymerase chain reaction (RT-PCR) for the identification of T.rubrum by comparing to DME and culture methods, from nail samples of patients with the complaints of onychomycosis. A total of 90 patients of whom 58 were male who were admitted to the dermatology outpatients clinics of our hospital with the complaints of color/shape changes in the nails and thickening of the nail, were included in the study, together with the 20 healthy volunteer subjects as controls. The nail scraping samples obtained from the patients and controls were examined with direct microscopy using 15% potassium hydroxide, dimethyl sulphoxide and chlorazole black mixture and cultivated onto Sabouraud dextrose agar with and without cycloheximide. For DNA isolation, after the disruption of nail samples with a steel tool, phenol-chloroform-isoamyl alcohol purification method were used. The amplification and demonstration of the T.rubrum DNA have been performed by using specific primers and probes following TaqMan protocol of RT-PCR (LightCycler-Roche, USA) method. Seventy-two of the patients yielded positive and 18 yielded negative results with DME. Growth of molds was detected in the cultures of 20 (27.8%) of the 72 DME positive patients and all of the isolates were identified as T.rubrum. No fungal growth was seen in the samples of 18 patients who were DME negative. In DME positive group, 67 (93%) patients were found to be positive in RT-PCR, while 8 (44.4%) patients were RT-PCR positive in DME negative group. All of the culture positive samples (n= 20) were also found positive in RT-PCR. All of the samples from the control group with healthy nails yielded negative results in DME, culture and RT-PCR methods. The performance of PCR method were compared to direct microscopy that had higher sensitivity than culture and the sensitivity, specificity, positive and negative predictive values of RTPCR assay were estimated as 93%, 56%, 89% and 67%, respectively. In conclusion RT-PCR was thought to be an efficient and rapid assay in the diagnosis of onichomycosis. Although RT-PCR seems more expensive than culture, for the centres which already have support for the molecular methods, the difference in total cost doesn't count much. In conclusion, by the use of molecular methods DNA isolation was successfully done from a relatively difficult clinical specimen, namely nail scraping, a protocole that could easily be applied in routine laboratory was established and species-level identification in a short time was accomplished in this study.

Investigation of fungi in massive nasal polyps: microscopy, culture, polymerase-chain reaction, and serology.

BACKGROUND: Although some clues exist about the causative relationship of fungi and chronic rhinosinusitis, the relationship of nasal polyps and fungi has not been enlightened. The purpose of this study was to determine whether a relationship exists between fungi and massive nasal polyps and to evaluate current available diagnostic techniques for detection of fungi. METHODS: Thirty cases of massive nasal polyposis (NP) were evaluated prospectively for fungal evidence and were compared with 18 cases of concha bullosa based on direct microscopy, fungal culture, serology, polymerase chain reaction (PCR), and sequencing. RESULTS: Fungal colonization was detected in 15 (50.0%) of the cases with massive NP, but only in 2 (11.1%) of the cases with concha bullosa. A significant difference was found between the study and the control groups in terms of fungal existence (p < 0.016). Direct microscopy was positive in 14 (46.7%) and 1 (5.6%) of the cases;fungal culture was positive in 8 (26.7%) and 4 (22.2%) of the cases; serology was positive in 9 (30.0%) and 2 (11.1%) of the cases; PCR was positive in 18 (60.0%) and 6 (33.3%) of the cases with massive NP and concha bullosa, respectively. CONCLUSION: Fungal colonization was found to be more common in massive NP patients compared with the control group. According to our results, microscopy and PCR were most sensitive techniques for detection of fungi.