Detection of Small Colony Variants Among Methicillin-Resistant Staphylococcus aureus Blood Isolates.

Staphylococcus aureus small colony variants (SCVs) are associated with chronic and persistent infections. Methicillin-resistant S. aureus (MRSA) SCVs cause more severe infections and mortality rates are higher in comparison with infections caused by MRSA. Our objective was to document the prevalence and phenotypical characteristics of SCVs among MRSA blood isolates. MRSA strains isolated from blood during 1999-2009 were evaluated retrospectively. Among 299 MRSA isolates, suspected colonies were inoculated onto Columbia blood agar and Schaedler agar. Columbia blood agar was incubated in normal atmosphere and Schaedler agar in 5-10% CO2, both at 35°C. If the small, nonpigmented, nonhemolytic colonies on Columbia blood agar were seen as normal-sized, hemolytic, and pigmented colonies on Schaedler agar, they were considered as MRSA SCVs. Six MRSA SCVs were detected. When subcultures were made, four of them reversed to phenotypically normal S. aureus, but two isolates were stable as SCV phenotype. The prevalence of SCVs among MRSA blood isolates was found as 6/299 (2%) with 2 (0.67%) stable. The detection of SCVs among MRSA blood isolates was reported from Turkey for the first time in this study. As the clinical significance of MRSA infections is well documented, evaluation of MRSA SCVs in clinical samples, especially from intensive care patients and those who have chronic and persistent infections are important to consider.

Association of clinical properties and compatibility of the QuantiFERON-TB Gold In-Tube test with the tuberculin skin test in patients with psoriasis.

BACKGROUND: Individuals with psoriasis show conflicting responses to the tuberculin skin test (TST), a commonly used screening test for latent tuberculosis infection. An alternative to TST is QuantiFERON-TB Gold In-Tube test (QFT-GIT), an in vitro interferon-gamma release assay. This study aimed to determine the effect of the clinical properties of psoriasis (disease severity and koebnerization status) on TST results and the agreement between the TST and QFT-GIT results in psoriatic patients. METHODS: One hundred patients with mild to severe psoriasis were enrolled in this prospective cross-sectional study. Psoriasis properties, including disease severity (psoriasis area and severity index score and koebnerization status), latent tuberculosis infection risk factors, and bacillus Calmette-Guérin vaccination history, were recorded. All patients underwent a TST and QFT-GIT. TST positivity cut-off point was ≥10 mm for bacillus Calmette-Guérin-vaccinated patients and ≥5 mm for non-vaccinated patients. RESULTS: Psoriasis area and severity index scores and koebnerization status did not correlate with TST diameters. Only one of the 23 koebnerization-positive patients developed koebnerization in response to TST. QFT-GIT positivity was prominently higher in the TST-positive group, and this was the only factor that differed between the TST-positive and TST-negative groups (P < 0.001). CONCLUSION: Tuberculin skin test results were not affected by psoriasis severity or koebnerization status. QFT-GIT positivity was prominently higher in the TST-positive group (P < 0.001). Overall agreement between TST and QFT-GIT results was moderate (κ = 0.413). Concurrent negativity (44%) was higher than concurrent positivity (27%).

Detection of the frequency of PER-1 type extended-spectrum ß-lactamase-producing Acinetobacter baumannii clinical isolates in Turkey: a multicenter study.

BACKGROUND/AIM: ß-Lactamases are an important resistance mechanism in Acinetobacter baumannii. Pseudomonas extended-resistance (PER-1) type ß-lactamase-producing strains have been reported from various geographic locations; however, PER-1 type ß-lactamases from Turkish hospitals have not been investigated extensively. The aim of this study was to determine the prevalence of PER-1 type ß-lactamases in A. baumannii isolates in various regions of Turkey. MATERIALS AND METHODS: A total of 763 clinical A. baumannii isolates were collected from 9 university hospitals and 2 state hospitals between 2008 and 2011. Molecular amplification of the OXA-51 gene from the A. baumannii genome was performed in order to verify identification of the species. Real-time polymerase chain reaction was used to detect blaPER-1 genes. RESULTS: PER-1 was detected in 24.6% of the isolates. The annual frequencies of the PER-1 enzyme were detected as 52.2%, 35.9%, and 8.3% in 2008, 2009, and 2010, respectively. PER-1 prevalence decreased gradually over time. The differences observed in PER-1 prevalence among the regions of Turkey were statistically significant (chi-square test; P < 0.001). CONCLUSION: These data demonstrate that the frequency of detection of PER-1 type ß-lactamases in A. baumannii species has decreased in Turkey. However, the increased carbapenem resistance, together with multidrug resistance, has created a worrisome situation regarding this pathogen.

Antimicrobial Susceptibilities of Clinical Acinetobacter baumannii Isolates With Different Genotypes.

BACKGROUND: The treatment of Acinetobacter baumannii infections is difficult. Carbapenems, sulbactam, and colistin are the most effective antibiotics. OBJECTIVES: The aim of this study was to evaluate the susceptibilities of genotypically different A. baumannii isolates to sulbactam, amikacin, netilmicin, meropenem, tigecycline and colistin. PATIENTS AND METHODS: Isolates from various clinical samples of patients with hospital-acquired infections that were identified by the VITEK 2 Compact system in our hospital's microbiology laboratory between January 2010 and March 2012 were included in the study. To determine genetic relatedness of the isolates, the rep-PCR method was used. The broth microdilution method was used for amikacin, netilmicin, meropenem and colistin, while E-test was used for sulbactam and tigecycline. RESULTS: Among the 300 isolates, 30 were found to be genotypically different and were evaluated in terms of their antimicrobial susceptibilities. All isolates were susceptible to colistin. The susceptibility rates were 66.6%, 50%, 36.6%, 30%, and 10% for netilmicin, tigecycline, sulbactam, amikacin, and meropenem, respectively. For carbapenem resistant isolates, the susceptibility rates were 66.6%, 51.8%, 33.3%, and 25.9% for netilmicin, tigecycline, sulbactam, and amikacin, respectively. The sulbactam minimum inhibitory concentration (MIC) 50 and MIC 90 were 8 µg/mL and 12 µg/mL, respectively. CONCLUSIONS: In this study, it was concluded that determining the cut-off value for MIC breakpoints for sulbactam alone has a critical impact on the susceptibility results.

Vancomycin and daptomycin minimum inhibitory concentration distribution and occurrence of heteroresistance among methicillin-resistant Staphylococcus aureus blood isolates in Turkey.

BACKGROUND: The aim of this study was to determine the distribution of vancomycin and daptomycin MICs among methicillin-resistant Staphylococcus aureus (MRSA) blood isolates, the prevalence of heterogeneous vancomycin-intermediate S. aureus (hVISA) and the relationship between hVISA and vancomycin MIC values. METHODS: A total of 175 MRSA blood isolates were collected from seven university hospitals in Turkey. All isolates were tested for susceptibility to vancomycin and daptomycin by reference broth microdilution (BMD) and by standard Etest method. BMD test was performed according to CLSI guidelines and Etest was performed according to the instructions of the manufacturer. All isolates were screened for the presence of the hVISA by using macro Etest (MET) and population analysis profile-area under the curve (PAP-AUC) methods. RESULTS: The vancomycin MIC50, MIC90 and MIC ranges were 1, 2, and 0.5-2 µg/ml, respectively, by both of BMD and Etest. The daptomycin MIC50, MIC90 and MIC ranges were 0.5, 1 and 0.125 -1 µg/ml by BMD and 0.25, 0.5 and 0.06-1 µg/ml by Etest, respectively. The vancomycin MIC for 40.6% (71/175) of the MRSA isolates tested was >1 µg/ml by BMD. No vancomycin and daptomycin resistance was found among MRSA isolates. Percent agreement of Etest MICs with BMD MICs within ±1 doubling dilution was 100% and 73.1% for vancomycin and daptomycin, respectively. The prevalence of hVISA among MRSA blood isolates was 13.7% (24/175) by PAP-AUC method. MET identified only 14 of the hVISA strains (sensitivity, 58.3%), and there were 12 strains identified as hVISA that were not subsequently confirmed by PAP-AUC (specificity, 92.1%). CONCLUSIONS: Agreement between BMD and Etest MICs is high both for vancomycin and daptomycin. Daptomycin was found to be highly active against MRSA isolates including hVISA. A considerable number of isolates are determined as hVISA among blood isolates. As it is impractical to use the reference method (PAP-AUC) for large numbers of isolates, laboratory methods for rapid and accurate identification of hVISA need to be developed.

Plasmodium falciparum malaria case originating from Uganda.

Malaria is the fifth infection leading to death in the world. Plasmodium species is the malarial parasite that infects human cells. The five species of the human Plasmodium parasites are P. falciparum, P. vivax, P. ovale, P. malariae and P. knowlesi. Recently, the World Health Organization reported that Uganda has the world's highest malaria incidence, with a rate of 478 cases per 1000 population per year. In this article, a patient who had specific clinical signs and symptoms of malaria after work-related travel to Uganda has been evaluated. The major clinical findings of the patient were chills and fever. After examination of thin and thick blood smears prepared from the peripheral blood of the patient, P. falciparum parasites were observed. Cerebral malaria was suspected as the patient's consciousness, orientation and cooperation had deteriorated. No Plasmodium was seen in control blood smears after treatment.

[Distribution of blaOXA genes in Acinetobacter baumannii strains: a multicenter study]. / Acinetobacter baumannii Izolatlarinda blaOXA Genlerinin Dagilimi: Çok Merkezli Bir Çalisma.

Acinetobacter baumannii is the most important agent of nosocomial infections within the Acinetobacter genus. This gram-negative coccobacillus is intrinsically resistant to many antibiotics used in antimicrobial therapy, and capable of developing resistance including carbapenems. The objective of this study was to develop a multiplex real time polymerase chain reaction (qPCR) kit for OXA subgroups in A.baumannii, and to investigate the distribution of OXA subgroups in A.baumannii strains isolated from geographically different regions of Turkey. A total of 834 A.baumannii clinical isolates collected from different state and university medical centers in 13 provinces (Afyonkarahisar, Ankara, Bolu, Elazig, Erzurum, Isparta, Istanbul, Kahramanmaras, Konya, Sakarya, Van) between 2008-2011, were included in the study. The isolates were identified by conventional methods and automated systems [Vitek2 (bioMerieux, ABD) and Phoenix (BD Diagnostic, MD)]. The susceptibility profiles of the isolates were studied with automated systems and standard disc diffusion method. All samples were subjected to qPCR to detect blaOXA-51-like, blaOXA-23-like and blaOXA-58-like genes. A conventional PCR method was also used to detect blaOXA-24-like gene. The resistance rates observed during the study period were as follows: 96.8% for amoxicillin-clavulanate, 86.8% for ciprofloxacin, 74.7% for gentamicin, 71.7% for amikacin, 73.5% for cefaperozone-sulbactam, 72.1% for imipenem and 73% for meropenem. Six hundred and two (72.2 %) isolates were resistant to both imipenem and meropenem. Colistin was found to be the most effective antibiotic against A.baumannii isolates with 100% susceptibility rate. All isolates were positive for blaOXA-51-like, however blaOXA-24-like gene could not be demonstrated in any isolate. Total positivity rates of blaOXA-23-like and blaOXA-58-like genes were found as 53.7% and 12.5%, respectively, while these rates were 74.4% and 17.3% in carbapenem-resistant isolates, respectively. Twenty-five isolates were positive for both blaOXA-23-like and blaOXA-58-like genes. All of the carbapenem-resistant isolates have OXA type genes with the exception of blaOXA-24-like gene. The positivity rates for blaOXA-23-like and blaOXA-58-like genes varied for each center. In addition, there was a decrease in the frequency of blaOXA-58-like gene, however both blaOXA-23-like gene and carbapenem resistance rates increased during the study period. In conclusion, high rates of resistance to carbapenems were also remarkable but A.baumannii strains keep on sensitivity to colistin. Both blaOXA-23-like and blaOXA-58-like genes were shown to be widespread in carbapenem-resistant A.baumannii clinical isolates. However, blaOXA-23-like gene positive strains were increased throughout the study. Currently, multiplex qPCR is the best way for rapid diagnosis of resistant bacteria for prevention of hospital-acquired infections. The multiplex qPCR kit developed in this study could be useful for rapid diagnosis and identify the frequencies of blaOXA-23-like, blaOXA-51-like and blaOXA-58-like genes in carbapenem-resistant A.baumannii clinical isolates.

[Evaluation of methicillin-resistant Staphylococcus aureus bacteremia and comparison of prognosis according to vancomycin MIC values: experience of the last ten years]. / Metisiline dirençli Staphylococcus aureus bakteriyemisinin incelenmesi ve vankomisin MIK degerlerine göre prognozun karsilastirilmasi: Son on yillik deneyim.

Hospital acquired methicillin-resistant Staphylococcus aureus (MRSA) infections are important health problems. Mortality and morbidity rates associated with MRSA infections are increasing with mortality rates being higher for MRSA bacteremia than the other clinical presentations of MRSA infections. Initiation of treatment immediately and use of appropriate antibacterial agents may lead to better clinical outcomes in MRSA bacteremia. The aims of this study were to evaluate the treatment and clinical outcomes of patients with MRSA bacteremia in a tertiary care hospital in Ankara, Turkey. Two hundred forty seven MRSA strains isolated from blood cultures at Hacettepe University Faculty of Medicine, Clinical Microbiology Laboratory between January 2000-December 2010, were evaluated retrospectively. Demographic characteristics, duration of bacteremia, types and duration of antibiotic treatment, presence of other pathogens and all other necessary information were collected from patients' registry. One hundred eighty four patients who had clinically significant bacteremia were analyzed. The mean age of the patients was 55 ± 17 years, of them 44.6% were female and 55.4% were male. The median length of hospital stay was 61 days. The median duration for the development of MRSA bacteremia from the time of admission was 23 days. Overall mortality rate was 54.9%, and mortality rate due to MRSA bacteremia was 19%. The rate of treatment success was 81%. There were 3 (1.6%) patients with vancomycin MIC value of 0.5 mg/L, 140 (76.1%) patients with 1 mg/L and 41 (22.3%) patients with 2 mg/L. The median duration from the time of MRSA bacteremia detection to the time of death was shorter in unsuccessfully treated group than successfully treated group (7 days vs. 30 days, p< 0.001). Thirty days mortality rate was higher in unsuccessfully treated group than successfully treated group (94.3% vs. 50.7%, p< 0.001). The median time interval between positive and negative cultures was 9.5 days. Number of patients with MRSA bacteremia had been decreasing for the last five years (36 patients in 2006, 18 in 2007, 16 in 2008, 12 in 2009 and one in 2010). In multivariate logistic regression analysis, it was shown that, intubation (OR: 5.086, 95% CI: 2.094-12.351; p< 0.001) and malignancy (OR: 2.789, 95% CI: 1.185-6.564; p= 0.019) were independent risk factors for mortality. In this study, it was shown that mortality rate was high in MRSA bacteremia and high MIC value was not an independent risk factor for mortality. It was also noted that when there was no clinical response to vancomycin, the therapy should be changed immediately. To decrease MRSA bacteremia rates in the hospital adherence to rules of infection control and prevention proved to be important factors.

Comparison of monolisa HCV Ag/Ab ULTRA with two anti-HCV assays for the detection of HCV infection in hospital setting.

In this study, we compared the performance of three serological assays (Monolisa HCV Ag/Ab ULTRA, Innotest HCV Ab IV enzyme immunoassay--EIA, and Ortho HCV 3.0 enzyme-linked immunosorbent assay--ELISA) for the detection of HCV infection. Ninety plasma samples were collected, representing 63 samples from groups at risk for acquiring HCV infection and 27 HCV RNA-positive samples. The results of Ortho HCV 3.0 ELISA, Innotest HCV Ab IV, and Monolisa HCV Ag/Ab ULTRA were fully concordant for 27 HCV RNA-positive samples. Ortho HCV 3.0 ELISA test and Innotest HCV Ab IV also gave the same results for risk groups, while three samples were found to be reactive by Monolisa HCV Ag/Ab ULTRA and were consequently found negative for HCV RNA. As two of the solely Monolisa HCV Ag/Ab ULTRA-positive samples were also hepatitis B s antigen (HBsAg)-positive, neutralization of HBsAg was performed but no arguments for the HBsAg interference were observed. In conclusion, the non-specific reactive signal was observed, in three samples using Monolisa HCV Ag/Ab ULTRA, to be negative by other serological assays, and observed to be negative in an HCV RNA assessment, a result that could not be attributed to the interference with HBsAg. In the context of diagnostic testing, no test for various HCV genotypes was observed to be superior to any other.

[Posttraumatic primary cutaneous aspergillosis with Candida guilliermondii infection in a healthy host]. / Saglikli bir olguda posttravmatik primer kütanöz aspergillozis ve Candida guilliermondii enfeksiyonu birlikteligi.

Opportunistic fungal infections are usually seen in immunocompromised patients. While Candida is the most prevalent agent in such infections, Aspergillus is at the second order. Primary cutaneous aspergillosis is most common in immunocompromised patients but can rarely be seen in healthy hosts as well. We report a case of posttraumatic primary cutaneous aspergillosis and Candida guilliermondii coinfection in a 70-years-old healthy man. The patient had an ulcerous lesion which developed in the site of a trauma on the middle finger of the right hand. Histopathological examination of the biopsy specimens revealed septate hyphae with dichotomous branching small circular blastospores. The cultures of the biopsy specimen yielded yellow-green colored, granular mold colonies and creamy white yeast colonies. Microscopic examination of the lactophenol cotton blue stained mold colonies indicated long conidiophores with vesicles surrounded by uniseriate phialides, compatible with Aspergillus flavus. Yeast colonies were identified as Candida guilliermondii by ID32C (BioMerieux, France) and by their microscopical morphology detected in corn meal-Tween 80 agar incubated at 25°C for 72 hours. The patient was treated properly with surgical debridement and itraconazole therapy. Since the immune system is compressed as a consequence of aging, cutaneous opportunistic fungal infections should be considered in the differential diagnosis of posttraumatic necrotic ulcers and black eschar in aged patients.

[Frequency of curable sexually transmitted infections among registered female sex-workers in Ankara City]. / Ankara ilinde kayitli seks calisani kadinlarda iyilestirilebilir cinsel yolla bulasan enfeksiyonlarin sikligi.

Sex-workers are considered as the high-risk population for sexually transmitted infections (STIs). Early diagnosis and treatment of curable STIs in this high-risk group have crucial importance in STI control and prevention of complications and transmission of infection. In this study, 146 registered female sex-workers in Ankara city were screened with rapid diagnostic tests (RDT) for causative agents of curable STIs such as, Neisseria gonorrhoeae, Chlamydia trachomatis, Trichomonas vaginalis and Treponema pallidum. To identify gonorrhoea, Gram stained endocervical smears were examined microscopically for the presence of gram-negative intracellular diplococci. For the diagnosis of chlamydial infection, an optic immunoassay (OIA) (Chlamydia OIA, Biostar, USA) as a RDT was performed by using endocervical specimens. For the detection of T. vaginalis, direct smears of vaginal swabs were examined for the presence of motile trophozoites first directly and after being cultured in Diamond's media for 24-48 hours of incubation. Syphilis was screened in the serum specimens by RPR (Omega, UK) test. There was no positive test results for gonorrhoea and syphilis however, the frequency of C. trachomatis and T. vaginalis in the study population was 1.4% and 0.7%, respectively. To provide comprehensive policies and optimal control strategies, a reliable source of data about the frequency and spectrum of STIs among high-risk populations and optimized effective screening programmes are required.

Molecular identification of adenoviral conjunctivitis in Turkey.

PURPOSE: The aim of the study was isolation of adenoviruses by cell culture and identification using polymerase chain reaction (PCR) and phylogenetic analyses in patients clinically diagnosed with viral conjunctivitis in Ankara, Turkey. METHODS: Conjunctival swabs from 34 patients with acute conjunctivitis were tested using cell culture isolation and PCR for adenovirus detection. PCR-positive samples were sequenced and typed. RESULTS: The positive results of adenovirus were 26.5% (9 of 34) by the PCR method and 20.6% by culture isolation. Nine samples positive at PCR were identified by phylogenetic analyses as human adenovirus 8 (HAdV-8) (4 of 9), HAdV-3 (3 of 9), HAdV-4 (1 of 9), and HAdV-B (1 of 9). CONCLUSIONS: Our study showed types of adenoviruses in patients with ocular infection that occurred in this region of Turkey for the first time. Furthermore, sequence-based typing method is an efficient, accurate, and rapid means of diagnosis and typing of the adenovirus and has significant clinical and epidemiologic implications. HAdV-8 was major type for acute conjunctivitis in Ankara, Turkey. Further studies are required to reveal the major types of HAdVs that cause ocular diseases in this region of the world.

Lens concentration of ofloxacin and lomefloxacin in an experimental endophthalmitis model.

UNLABELLED: Abstract. BACKGROUND: Bacterial endophthalmitis is a serious complication of ocular surgery and penetrating trauma. The primary causative organisms are strains of Staphylococcus aureus and Staphylococcus epidermidis. Fluoroquinolones are widely used to treat endophthalmitis. There are a few studies on the penetration of fluoroquinolones into the lens in inflamed eyes. A literature search did not identify any data regarding penetration of topical ofloxacin into the lens in normal and inflamed eyes. OBJECTIVE: The aim of this study was to determine the penetration of topical ofloxacin and lomefloxacin into the lens in a rabbit endophthalmitis model. METHODS: New Zealand white rabbits were randomly divided into 2 groups. The left eyes were infected with an intravitreal inoculation of S aureus. The right eyes were used as a noninoculated control. Groups 1 and 2 received topical ofloxacin and lomefloxacin treatment, respectively, 24 hours after the inoculation. Two drops of the study drugs were instilled in the eyes every 30 minutes for 3 hours and then every 60 minutes for 3 hours. Lens samples were obtained 30 minutes after the last ofloxacin or lomefloxacin drops were administered. High-performance liquid chromatography was used to determine the fluoroquinolone concentration. RESULTS: Ten rabbits were equally divided into the 2 treatment groups. There was no significant difference in mean (SD) lens concentrations between the control and inoculated eyes in either treatment group-ofloxacin (0.26 [0.32] µg/mL vs 0.11 [0.05] µg/mL, respectively) and lomefloxacin (0.50 [0.87] µg/mL vs 0.12 [0.08] µg/mL, respectively). CONCLUSION: The results of this small experimental study found that topical ofloxacin and lomefloxacin can accumulate in the crystalline lens after installation. Inflammation did not affect the penetration of ofloxacin or lomefloxacin into the lens.