Prevalence and genetic diversity of Staphylococcus aureus small-colony variants in cystic fibrosis patients.

Staphylococcus aureus small-colony variants (SCVs) are being isolated more frequently in cystic fibrosis (CF) patients. We aimed to determine the prevalence of S. aureus SCVs and their phenotypic and genotypic properties in CF patients admitted to a university hospital. Specimens of 248 patients were examined during a period of 11 months. Colonies supposed to be SCVs were evaluated on Columbia blood agar, mannitol salt agar, and brain-heart infusion agar with 5% NaCl (BHIA 5% NaCl). Strains were confirmed by S. aureus nucA PCR. Antibiotic susceptibilities of SCVs and simultaneously isolated S. aureus strains were determined for oxacillin, gentamicin, trimethoprim-sulphamethoxazole, vancomycin, ciprofloxacin, linezolid, and tigecycline. Genetic relatedness between SCVs and normal S. aureus strains was determined with a pulsed-field gel electrophoresis (PFGE) method. S. aureus SCVs were detected in 20 of 248 patients (8.1%). The highest SCV isolation rate was obtained with MSA, followed by BHIA 5% NaCl. Auxotrophism for thymidine was demonstrated in six SCVs. The tigecycline susceptibilities of 48 SCV strains isolated in this study showed higher MIC values than those of 33 simultaneously isolated normal S. aureus strains. Whereas SCVs and normal S. aureus strains showed identical genotypes in 14 of the patients, five patients showed different genotypes. This first study from Turkey evaluating S. aureus SCVs in CF patients has indicated the importance of considering and reporting SCVs in chronic infections such as CF. The presence of SCVs will probably indicate persistent infection, and this might impact on antibiotic treatment decisions, as they are more resistant to antibiotics.

Clinical value of cystatin c determination in children with lymphoma.

Cystatin, a proteinase inhibitor, is involved in the intracellular catabolism of proteins. We investigated the change in concentration of serum Cystatin C (CysC) in children with lymphomas and its diagnostic utility. Twenty-eight newly diagnosed patients with lymphoma were included in this study. The male/female ratio was 20/8, with a median age of 8.5 years (range 3-17 years). Thirteen patients had Hodgkin's lymphoma (HL) and 15 had non-Hodgkin's lymphoma (NHL). Cystatin C concentration was determined at the time of diagnosis and during remission. In the entire group, CysC concentrations at diagnosis and during remission were 0.87+/-0.29 mg/L and 0.86+/-0.21 mg/L, respectively (p=0.93). In the NHL group, CysC concentrations at diagnosis and remission were 0.89+/-0.32 mg/L and 0.85+/-0.23 mg/L, respectively (p=0.73). The CysC concentrations in the HL group at diagnosis and remission were 0.88+/-0.36 mg/L and 0.88+/-0.18 mg/L, respectively (p=0.73). No significant difference was observed between CysC concentrations in the HL (0.88+/-0.36 mg/L) and NHL (0.89+/-0.32 mg/L) groups. Cystatin C concentrations in all the patients with localized versus those with advanced disease were 0.91+/-0.41 mg/L and 0.88+/-0.3 mg/L, respectively (p=0.83). Cystatin C concentrations of the patients with localized and advanced HL were 0.95+/-0.45 mg/L and 0.77+/-0.14 mg/L, respectively, (p=0.41). Cystatin C level was higher in patients with localized disease, in those without B symptoms, and, at diagnosis, in those with an unfavorable response (Tab. 2, Ref. 20). Full Text in free PDF www.bmj.sk.

Sandfly fever virus activity in central/northern Anatolia, Turkey: first report of Toscana virus infections.

Sandfly fever viruses (SFVs) cause febrile diseases as well as aseptic meningitis/encephalitis and include serotypes sandfly fever Sicilian virus (SFSV), sandfly fever Naples virus (SFNV) and Toscana virus (TOSV). Infections are endemic in the Mediterranean basin and data on SFV activity in Turkey are limited. In this study, sera from 1533 blood donors from the Ankara, Konya, Eskisehir and Zonguldak provinces of Turkey were evaluated for SFV exposure by indirect immunofluorescence test (IIFT) and confirmed by virus neutralization test (VNT). One hundred and two patients with central nervous system (CNS) infections of unknown aetiology were also tested via IIFT and real-time reverse-transcription PCR for SFV/TOSV. Rate of overall IgG reactivity in IIFT was 32.9% (505/1533) among blood donors. TOSV exposure was confirmed by VNT in all study regions. Exposure to the recently-identified serotype sandfly fever Turkish virus, as evaluated by VNT, was revealed in Konya and Ankara. SFNV exposure was identified in Konya and SFSV was observed to be present in all regions except Zonguldak. TOSV RNA was detected in 15.7% (16/102) and was accompanied by TOSV IgM in 25% (4/16) of the patients. Partial L and S sequences suggested that TOSV circulating in Turkey can be grouped into TOSV genotype A strains. Exposure to TOSV and other SFV serotypes was revealed in blood donors and CNS infections by TOSV were identified for the first time in Turkey. Infections are observed to be endemic in central Anatolia and should be considered as aetiologic agents in cases/outbreaks of fever and meningoencephalitis.

Confirmed exposure to tick-borne encephalitis virus and probable human cases of tick-borne encephalitis in Central/Northern Anatolia, Turkey.

Tick-borne encephalitis virus (TBEV) is the aetiological agent of tick-borne encephalitis (TBE), a potentially fatal central nervous system infection of humans. TBE is endemic in many areas of Europe and Asia; however, very scarce data on TBEV activity are available from Turkey. We aimed to identify TBEV exposure in healthy blood donors and the impact of TBEV in central nervous system infections in Central/Northern Anatolia. Two-thousand four hundred and fifty four sera, collected from blood donors at Ankara, Konya, Eskisehir and Zonguldak branches of the Turkish Red Crescent Middle Anatolia Regional Blood Center, were analysed for TBEV serosurveillance. Paired serum and cerebrospinal fluid samples from 108 patients with the diagnosis of aseptic meningitis/encephalitis of unknown aetiology were also evaluated to identify TBE and neuroborreliosis cases. Commercial enzyme-linked immunosorbent assays and indirect immunofluorescence tests were employed for antibody detection. Forty-seven donor samples (1.9%) were reactive for TBEV IgG. In 25 persons with IgG reactivity (53.1%), risk factors for tick-borne infections were revealed. One sample from Zonguldak province (1/198; 0.5%) in the Black Sea region of Turkey was confirmed to possess neutralizing antibodies via plaque reduction neutralization test. TBEV IgM was detected in 9.2% (8/108) of the patients. IgM was accompanied by IgG reactivity in two persons where, in one, recent history of a tick bite was also identified. Intrathecal antibody production for TBEV could not be demonstrated. No evidence for Borrelia infections could be found. Confirmed exposure to TBEV and/or an antigenically similar tick-borne flavivirus is documented for the first time in blood donors in Zonguldak in Northern Anatolia. Probable cases of TBE have also been identified from Central Anatolia. The epidemiology of TBEV activity in Turkey needs to be assessed and benefits of vaccination for general population, risk groups or travellers must be considered.

Antimicrobial resistance in gram-negative hospital isolates: results of the Turkish HITIT-2 Surveillance Study of 2007.

Resistance rates to amikacin, ciprofloxacin, ceftazidime, cefepime, imipenem, cefoperazone/sulbactam and piperacillin/tazobactam in Escherichia coli (n= 438), Klebsiella pneumoniae (n= 444), Pseudomonas aeruginosa (n= 210) and Acinetobacter baumanni (n=200) were determined with e-test in a multicenter surveillance study (Hitit-2) in 2007. ESBL production in Escherichia coli and K. pneumoniae was investigated following the CLSI guidelines. Overall 42.0% of E.coli and 41.4% of K. pneumoniae were ESBL producers. In E. coli , resistance to imipenem was not observed, resistance to ciprofloxacin and amikacin was 58.0% and 5.5% respectively. In K. pneumoniae resistance to imipenem, ciprofloxacin and amikacin was 3.1%, 17.8% 12.4% respectively. In P. aeruginosa the lowest rate of resistance was observed with piperacillin/tazobactam (18.1%). A. baumanni isolates were highly resistant to all the antimicrobial agents, the lowest level of resistance was observed against cefoperazone/sulbactam (52.0%) followed by imipenem (55.5%). this study showed that resistance rates to antimicrobials are high in nosocomial isolates and show variations among the centers.

Efficacy of nasopharyngeal culture in identification of pathogen in middle ear fluid in chronic otitis media with effusion.

PURPOSE: Chronic otitis media with effusion (OME) is the leading cause of hearing loss during childhood. In bacterial etiology of OME, the most frequent pathogens responsible are Haemophilus influenzae followed by Streptococcus pneumoniae and Moraxella catarrhalis. This study aimed at evaluating the accuracy of nasopharyngeal (NP) specimens in the identification of pathogens in the middle ear fluid (MEF) in patients with OME. MATERIALS AND METHODS: In this cross sectional, case-control study, 95 MEFs and 53 NP secretion specimens were obtained from 53 children. As a control group, 102 NP specimens were taken from children having an operation other than an otological disease. Conventional culture methods and multiplex-PCR method have been used to determine the etiology of OME; NP carriage between cases and control groups were compared using conventional culture methods. Pearson Chi-Square and Fisher's Exact tests were used in statistical analysis. RESULTS: Bacteria were isolated by culture in 37.9% of MEF specimens, 14.7% of which belonged to the group H. influenzae, S. pneumoniae and M. catarrhalis. PCR was positive in 30.5% specimens targeting the same pathogens. There was a two-fold increase in carriage rate of S. pneumoniae and H. influenzae in patients than controls for each pathogen. CONCLUSION: PCR is a more reliable method to detect middle ear pathogens in MEF in comparison with the conventional culture methods. The NP colonization wasn't found to be an indicator of the pathogen in MEF although middle ear pathogens colonize more in nasopharynx of diseased children.

Simultaneous triple organ specific autoantibody profiling in adult patients with type 1 diabetes mellitus and their first-degree relatives.

AIMS: We aimed to document prevalence and clinical presentations of seropositivities for glutamate decarboxylase (GAD)-antibody, celiac's disease (CD) and autoimmune thyroiditis (AIT) in adult patients with type 1 diabetes mellitus (T1DM), and their first-degree relatives. METHODS: Sixty-five patients with T1DM, 124 first-degree relatives and 65 healthy controls were screened for GAD-antibody, anti-thyroid peroxidase (ATPO), anti-thyroid stimulating hormone receptor (TSHR), anti-tissue transglutaminase and anti-gliadin antibodies in a matched case-control study. RESULTS: Prevalence of more than one seropositivity for CD-associated antibodies in T1DM-group is 6.0 times increased, compared with controls (p < 0.05). ATPO seropositivity is 5.3 times increased in T1DM group (p < 0.05), but TSHR antibody is comparable with controls (p > 0.05). Seropositivities for T1DM, AIT and CD are 4.3, 1.9 and 2.4 times more prevalent among first-degree relatives respectively, compared with controls (p < 0.05). Pathologically confirmed cases with CD among first-degree relatives were all identified at screening. In contrast, all of pathologically confirmed cases with CD in T1DM group, were either previously diagnosed or symptomatic at time of screening. In the group of patients with T1DM, 31% of seropositive cases for anti-ATPO were clinically latent for AIT, and 74% of ATPO (+) cases were identified at current screening study. Sixty-four per cent of ATPO (+) first-degree relatives were clinically latent for AIT, and 54% were identified at screening. CONCLUSION: Type 1 diabetes mellitus, CD and AIT represent a significant overlap in an adult population with already-diagnosed T1DM and their first-degree relatives. With regard to clinical presentations, CD was less likely to be clinically silent than AIT among patients with T1DM.

A comparison of PCR detection of mecA with oxacillin disk susceptibility testing in different media and sceptor automated system for both Staphylococcus aureus and coagulase-negative staphylococci isolates.

PURPOSE: To evaluate three methods for 406 isolates of Staphylococcus aureus and coagulase-negative staphylococci (CNS) for the detection of methicillin resistance (MR) using National Committee for Clinical Laboratory Standards (NCCLS) new interpretive criteria. METHODS: We used polymerase chain reaction (PCR) as a gold standard method to evaluate three methods [disk diffusion with Mueller-Hinton agar (MHA) and mannitol salt agar (MSA) and Sceptor system (Becton Dickinson, USA)] for the detection of mecA gene. The isolates that were methicillin-resistant with any of the three tests were evaluated further for MR by E-test. RESULTS: MHA, MSA and Sceptor showed sensitivities of 100, 100 and 99% for S. aureus and 100, 82.6 and 72.1% for CNS, respectively. The specificities of the same methods were found as 100, 90.1 and 99.3% for S. aureus and 79.2, 95.8 and 97.2% for CNS, respectively. E-test showed 100% sensitivity for both S. aureus and CNS. Forty-eight CNS and 16 S. aureus isolates, which presented discrepancies with the three phenotypic methods (MHA disk diffusion method, MSA disk diffusion method and Sceptor), were correctly classified as resistant/susceptible with the E-test when compared with PCR. Only five CNS isolates, which were mecA-negative with PCR were resistant with E-test. Analysis of 248 S. aureus revealed that MHA is superior to other phenotype-based susceptibility testing methods in detecting MR. When we examined the results of 158 CNS, none of the three methods proved efficient in detecting MR. CONCLUSIONS: We conclude that although the accuracy of the MHA disk diffusion test for the detection of MR approaches the accuracy of PCR for S. aureus isolates, the need for easy and reliable methods of detecting MR in CNS still remains.

Use of plasma Epstein-Barr virus DNA monitoring as a tumor marker in follow-up of patients with nasopharyngeal carcinoma: preliminary results and report of two cases.

Recent studies suggest that plasma Epstein-Barr virus (EBV) DNA may reflect tumor burden in patients with nasopharyngeal cancer. A prospective study was initiated to investigate this correlation in 125 patients (34 pretreatment [Group A], 78 in remission [Group B] and 13 relapsed [Group C]) and 19 healthy controls. In group A, EBV DNA was detected in plasma samples of 24 (70%) patients. In Group B, EBV DNA was detected in 7 patients (range 77-13,731 copies/mL) and further imaging in all but one of these patients revealed active disease confirmed by ultrasound-guided fine-needle biopsy. There was only one false-positive case; this patient is currently under follow-up. Here we describe 2 of the 7 patients with detectable plasma EBV DNA in whom recurrence was documented by PET scan during follow-up. Our results showed that in group B the positive predictive value of quantitative analysis of plasma EBV DNA was 85%. Quantitative analysis of EBV DNA in plasma seems to become an integral part of screening, staging, monitoring, and prediction of relapse in patients with nasopharyngeal carcinoma. However, previous studies cannot be considered definitive and more reports on the use of this technique are urgently needed from both endemic and non-endemic regions.

Varicella vaccination in children with lymphoma and solid tumours.

BACKGROUND: Varicella infection can be a severe disease, especially in immunosuppressed patients. Here, experience with live varicella vaccine to prevent varicella infection is reported in children who were undergoing treatment for lymphoma and solid tumours. METHODS: 40 children, aged between 12 months and 15 years with no clinical history of varicella, were vaccinated with live varicella vaccine. All received two doses of the vaccine subcutaneously 4 weeks apart. Serum samples were taken before the first dose and 6 weeks after the second dose of vaccine. RESULTS: Before vaccination, 32 patients were seronegative for varicella and eight were seropositive. Seroconversion was observed 6 weeks after the second dose in 24 of the 32 (75%) seronegative children. In 4 of 8 previously seropositive patients, antibody titres increased after immunisation. Zoster infection occurred 5 weeks after the second dose of vaccine in only one previously seronegative child. 7 children, who had responded to the vaccine, have been exposed to varicella in their families or in school without contracting clinical disease. CONCLUSION: Although the small number of patients in our group prevents us from drawing definitive conclusions, the varicella vaccine seems to be well tolerated and can be administered to children with lymphoma and solid tumours undergoing treatment.

Tonsillar abscess formation due to herpes simplex type-1 in a severely immunocompromised stem cell transplant patient with chronic myeloid leukemia.

Herpes simplex virus (HSV) causes life-threatening infections in immunocompromised patients such as transplant recipients and patients with hematologic malignancies. We herein describe the case of a patient with chronic myeloid leukemia blastic transformation who developed severe herpetic tonsillitis complicated by tonsillar abscess formation. Abscess formation was determined by computed tomography, whereas tonsillitis due to HSV was confirmed by pathologic and immunohistochemical examinations of the tonsillar biopsy. For molecular confirmation, HSV DNA was amplified by LightCycler PCR and type (HSV-1) determined by melting point analysis. The patient responded promptly to antiviral treatment and there were no signs of recurrent infection at the follow-up. To our knowledge, this case is unique for being the first case of tonsillar abscess formation due to HSV-1, also emphasizing the importance of herpetic infections in the differential diagnosis of oropharyngeal small-sized lesions in the immunocompromised patient population.

Stenotrophomonas maltophilia: antimicrobial resistance and molecular typing of an emerging pathogen in a Turkish university hospital.

Despite its limited pathogenicity, Stenotrophomonas maltophilia is an emerging nosocomial pathogen. This study investigated the isolation frequency, antimicrobial resistance and genotypic relationships of 205 S. maltophilia isolates from 188 patients in a university hospital between 1998 and 2003. Susceptibility profiles for 11 antimicrobial agents were determined by the NCCLS agar dilution method for non-fermentative bacteria, while enterobacterial repetitive intergenic consensus sequence (ERIC)-PCR and pulsed-field gel electrophoresis (PFGE) were used for genotyping of the isolates. Of the 205 isolates, 56.1% were isolated in the last 2 years of the study. The risk of S. maltophilia isolation was higher in intensive care units, S. maltophilia was isolated mostly (86.8%) after hospitalisation for >or= 48 h, and 90.4% of the patients had underlying diseases. Resistance levels were>60% for all antimicrobial agents tested except co-trimoxazole. High genetic diversity was found among the S. maltophilia isolates, and cross-infection with S. maltophilia was not common. Although ERIC-PCR revealed fewer genotypes than PFGE, it proved to be a rapid and easy method for S. maltophilia genotyping, and was more economical than PFGE.

Antimicrobial resistance of Salmonella enterica group C strains isolated from humans in Turkey, 2000-2002.

Fifty-three Salmonella enterica group C isolates obtained from various human samples (47 stool, 4 blood and 2 urine) in ten provinces of Turkey between 1 July 2000 and 30 June 2002 were serotyped and resistance to antimicrobials was investigated by agar dilution tests. The isolates were identified as S. Choleraesuis (11), S. Hadar (7), S. Irumu (4), S. Virchow (3), S. Tallahassee (3), S. Paratyphi C (2), S. Braenderup (2), S. Othmarschen (2), S. Menston (2), S. Concord (2), S. Infantis (2), S. Kottbus (2), S. Edinburg (1), S. Oranienburg (1), S. Muenchen (1) and S. Malmoe (1). Antimicrobial resistance rates of S. enterica groups C1 and C2 were high for ampicillin (26% and 60%, respectively), amoxicillin/clavulanic acid (11% and 40%), chloramphenicol (16% and 27%) and tetracycline (3% and 40%). The percentages of strains sensitive to all antimicrobials were 58% and 33%, respectively. Multiresistance was not observed in group C1 isolates, but the rate of multiresistant isolates was 13% in group C2. The rate of decreased ciprofloxacin susceptibility (CipL) was 61% in serogroup C1 and 20% in serogroup C2. These results indicated that S. enterica group C infections in humans were not infrequent in Turkey and that multiple antimicrobial resistance was common within these strains.

Antimicrobial resistance patterns and serotype distribution among Salmonella enterica strains in Turkey, 2000-2002.

Since Turkey currently lacks a national reference center for Salmonella infections, the present study was conducted to document the distribution of serotypes and antimicrobial resistance patterns among Salmonella enterica isolates recovered from clinical samples in ten Turkish provinces over a 2-year period. Among the 620 Salmonella enterica isolates recovered between 1 July 2000 and 30 June 2002, strains belonging to the serotypes Enteritidis (47.7%), Typhimurium (34.7%), Paratyphi B (6.0%), Typhi (2.9%), Paratyphi A (0.2%) and serogroup C (8.5%) were found. Resistance to multiple antimicrobial agents was particularly high among Salmonella Typhimurium isolates (76.7%), and resistance or decreased susceptibility to ciprofloxacin (MIC> or =0.125 mg/l) was demonstrated in Salmonella Paratyphi B, Salmonella Typhimurium and Salmonella Enteritidis strains. All of the Salmonella Typhi isolates were susceptible to ciprofloxacin. The results indicate that decreased susceptibility to ciprofloxacin is an emerging problem in Salmonella enterica in Turkey, particularly in multiresistant strains.

Seroprevalence and some correlates of Helicobacter pylori at adult ages in Gülveren Health District, Ankara, Turkey.

The purpose of the study was to detect the frequency and distribution of Helicobacter pylori in the Gülveren Health Centre service area among residents aged between 25 and 64 years and to evaluate the relation of H. pylori infections with general health status, socioeconomic status, and some lifestyle habits. The study included a representative sample of Gülveren Health Centre residents, aged between 25 and 64 years. A stratified random sample of 1672 individuals was selected for study purposes out of 10,569 residents, stratified by age and gender. A standardized questionnaire was completed for all study participants using a face-to-face interview and all participants were invited to the local health centre for a thorough physical examination and blood tests. The overall prevalence of H. pylori was found to be 77.5 % among individuals aged between 25 and 64 years. The frequency of H. pylori was higher among individuals with low socioeconomic status; those who migrated to Ankara after the age of 20 years; members of large families (household size of 4 or above); non-alcohol drinkers; and those who regularly drink tea.

Non tuberculous mycobacteria (NTM) in patients with underlying diseases: results obtained by using polymerase chain reaction-restriction enzyme analysis between 1997-2002.

In this study, we aimed to evaluate the frequency of non-tuberculous mycobacteria (NTM) isolated from clinical specimens using Polymerase Chain Reaction-Restriction Enzyme Analysis (PCR-REA) and to investigate the patients who had clinically significant NTM infections in our hospital through the five year period from May 1997 to June 2002. A total of 364 mycobacterial strains isolated from clinical specimens which gave positive growth index in the BACTEC 460 radiometric system in Hacettepe University Hospital Clinical Microbiology Laboratory were evaluated by PCR-REA and clinical data were obtained from the patient records. Three hundred and one of the strains (82.7%) were identified as Mycobacterium tuberculosis and 63 (17.3%) were identified as nontuberculous mycobacteria. Seven (11.1%) of 63 NTM patients were regarded as having clinical mycobacteriosis. Chronic obstructive pulmonary disease and other pre-existing lung diseases were seen in 39 (61.9%) of the patients, 11 (17.5%) of'the patients had chronic renal failure. Four (6.3%) and 9 (14.3%) of them had AIDS and carcinomas, respectively. PCR-REA was found to be a reliable method for typing of our mycobacterial isolates to the species level. These data may shed light on the epidemiology of the mycobacterial species and help to select a proper treatment regimen.

In vitro activities of new quinolones against Brucella melitensis isolated in a tertiary-care hospital in Turkey.

We have evaluated the in vitro activities of seven fluoroquinolones against 69 strains of Brucella melitensis. According to their minimum inhibitory concentration for 90% growth (MIC(90)) values, the most active agent was found to be sparfloxacin (MIC(90) 0.12 mg/L) followed by levofloxacin, ciprofloxacin, ofloxacin (MIC(90) 0.50 mg/L) and grepafloxacin (MIC(90) 1 mg/L), gemifloxacin (MIC(90) 2 mg/L) and gatifloxacin (MIC(90) 4 mg/L).