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1.
Ann Med ; 55(1): 266-275, 2023 12.
Article in English | MEDLINE | ID: mdl-36579976

ABSTRACT

OBJECTIVE: To evaluate the serotype distribution and antibiotic resistance in pneumococcal infections in adults and to provide a perspective regarding serotype coverage of both current and future pneumococcal vaccines. PATIENTS AND METHODS: This passive surveillance study was conducted with the Streptococcus pneumoniae strains isolated from the specimens of patients with pneumonia (materials isolated from bronchoalveolar lavage), bacteraemia, meningitis, pleuritis and peritonitis between 2015 and 2018. Serogrouping and serotyping were performed by latex particle agglutination and by conventional Quellung reaction using commercial type-specific antisera, respectively. The strains were analysed for penicillin, cefotaxime, erythromycin and moxifloxacin susceptibilities by E-test. RESULTS: In the whole study group (410 samples from adults aged ≥18 years), the most frequent serotypes were 3 (14.1%), 19 F (12%) and 1 (9.3%). The vaccine coverage for PCV13, PCV15, PCV20 and PPV23 was 63.9%, 66.6%, 74.1% and 75.9%, respectively, in all isolates. Penicillin non-susceptibility in invasive pneumococcal disease (IPD) was 70.8% and 57.1% in the patients aged <65 and ≥65 years, respectively. About 21.1% and 4.3% of the patients with and without IPD had cefotaxime resistance. Non-susceptibility to erythromycin and moxifloxacin was 38.2% and 1.2%, respectively. CONCLUSIONS: The results revealed that novel PCV vaccines may provide improved coverage as compared with the currently available vaccine, PCV13. The significant antibiotic resistance rates imply the need to extend the serotype coverage of the vaccines. Continuing the surveillance in pneumococcal diseases is critical to explore the serotype distribution and incidence changes of IPD cases in the population and to inform policy makers to make necessary improvements in the national immunization programmes.Key messagesThis multicentre study demonstrated the most recent serotype distribution and antibiotic resistance in adult population in Turkey.Shifting from PCV13 to novel conjugated vaccines will significantly increase the coverage.Continuing the surveillance in pneumococcal diseases is critical to explore the serotype distribution changes and the incidence of cases with invasive pneumococcal disease in the population.


Subject(s)
Pneumococcal Infections , Streptococcus pneumoniae , Adult , Humans , Infant , Adolescent , Serogroup , Pneumococcal Vaccines , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/therapeutic use , Moxifloxacin , Turkey/epidemiology , Pneumococcal Infections/epidemiology , Pneumococcal Infections/prevention & control , Pneumococcal Infections/drug therapy , Cefotaxime/pharmacology , Cefotaxime/therapeutic use , Erythromycin , Penicillins/pharmacology , Penicillins/therapeutic use
2.
Pediatr Infect Dis J ; 37(4): 373-375, 2018 04.
Article in English | MEDLINE | ID: mdl-29189676

ABSTRACT

Cryptococcosis is less common in children than in adults but remains an important cause of pneumonia and meningoencephalitis in both immunocompromised and immunocompetent patients. Intracranial hypertension commonly complicates cryptococcal meningitis and may cause significant visual and neurologic morbidity and mortality. Early and aggressive management of intracranial hypertension in accordance with established guidelines reduces the risk of long-term complications and death. In this case report, we present a 12-year-old girl with cryptococcal meningitis, pneumonitis and dermatitis complicated with cranial nerve palsy and loss of vision. She was successfully treated with serial cerebrospinal fluid drainage, antifungal and interferon gamma therapy.


Subject(s)
Cranial Nerve Diseases/etiology , Cranial Nerve Diseases/pathology , Intracranial Hypertension/etiology , Intracranial Hypertension/pathology , Meningitis, Cryptococcal/diagnosis , Meningitis, Cryptococcal/pathology , Antifungal Agents/administration & dosage , Blindness/etiology , Blindness/pathology , Child , Dermatitis/etiology , Dermatitis/pathology , Drainage , Female , Humans , Interferon-gamma/administration & dosage , Intracranial Hypertension/complications , Meningitis, Cryptococcal/complications , Meningitis, Cryptococcal/therapy , Pneumonia/etiology , Pneumonia/pathology , Treatment Outcome
3.
Clin Lab ; 63(4): 699-703, 2017 04 01.
Article in English | MEDLINE | ID: mdl-28397475

ABSTRACT

BACKGROUND: Infections due to Candida species are major causes of morbidity and mortality in humans, causing a diverse spectrum of clinical disease ranging from superficial and mucosal infections to invasive disease. Several authors have demonstrated that mortality is closely linked to both timing of therapy and/or source control. The rapid identification of pathogenic species is helpful to start timely and effective antifungal therapy. The aim of this study was to assess the performance of the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) system for the correct and rapid identification of yeast isolates causing bloodstream infection. METHODS: Between January 2014 and January 2015, a total of 117 yeast like organisms isolated from blood culture samples of 117 episodes from 102 patients who had blood stream infections were included in the study. The isolates were identified by MALDI-TOF MS. The results were compared with those obtained by the standard mycological methods and/or sequence analysis. RESULTS: One hundred and seventeen yeast isolates including 115 Candida spp and two non-Candida yeasts were analysed. The Biotyper correctly identified 115 (98.3%) isolates to the genus level and 102 (87.2%) isolates to the species level using the manufacturer's recommended cutoff scores. CONCLUSIONS: The Bruker Biotyper is a rapid, easy, inexpensive, and highly reliable system for the identification of yeast isolates. Early identification with MALDI-TOF MS would save time for determination of antifungal susceptibility and proper treatment strategy. The expansion of the database of the library by addition of less common species will improve the performance of the system.


Subject(s)
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Antifungal Agents , Bacteremia , Candida , Humans , Saccharomyces cerevisiae
4.
Mikrobiyol Bul ; 50(1): 21-33, 2016 Jan.
Article in Turkish | MEDLINE | ID: mdl-27058326

ABSTRACT

Carbapenems are the choice of treatment in infections caused by multidrug resistant Enterobacteriaceae. In recent years carbapenem-resistant Enterobacteriaceae isolates due to carbapenemases have been increasingly reported worldwide. Multicenter studies on carbapenemases are scarce in Turkey. The aim of this study was to determine the distribution of carbapenemases from different parts of Turkey as a part of the European Survey of Carbapenemase Producing Enterobacteriaceae (EuSCAPE) project. Beginning in November 2013, carbapenem-resistant isolates resistant to at least one of the agents, namely imipenem, meropenem, and ertapenem were sent to the coordinating center. Minimum inhibitory concentrations for these carbapenems were determined by microdilution tests following EUCAST guidelines. Production of carbapenemase was confirmed by combination disk synergy tests. Types of carbapenemases were investigated using specific primers for VIM, IMP; NDM, KPC and OXA-48 genes by multiplex polymerase chain reaction. In a six month period, 155 suspected carbapenemase-positive isolates were sent to the coordinating center of which 21 (13.5%) were E.coli and 134 (86.5%) were K.pneumoniae. Nineteen (90.5%) strains among E.coli and 124 (92.5%) strains among K.pneumoniae were shown to harbour at least one carbapenemase gene by molecular tests, with a total of 92.3% (143/155). Carbapenemases were determined as a single enzyme in 136 strains (OXA-48: 84.6%; NDM: 6.3%; VIM: 2.8%; IMP: 1.4%) and as a combination in seven isolates (OXA-48 + NDM: 2.1%; OXA-48 + VIM: 2.1%; VIM + NDM: 0.7%). KPC was not detected in any of the isolates. According to the microdilution test results, resistance to imipenem, meropenem and ertapenem in OXA-48 isolates were 59.5%, 52.9% and 100%, respectively. The combination disk synergy test was 100% compatible with the molecular test results. As most of the OXA-48 producing isolates were susceptible to meropenem but all were resistant to ertapenem, ertapenem seems to be the most sensitive agent in screening carbapenemases in areas where OXA-48 is prevalent and phenotypic combination tests can be useful in centers where molecular tests are not available.


Subject(s)
Anti-Bacterial Agents/pharmacology , Bacterial Proteins/metabolism , Carbapenems/pharmacology , Escherichia coli/enzymology , Klebsiella pneumoniae/enzymology , beta-Lactamases/metabolism , Bacterial Proteins/genetics , Drug Resistance, Multiple, Bacterial/genetics , Ertapenem , Escherichia coli/drug effects , Escherichia coli Proteins/genetics , Escherichia coli Proteins/metabolism , Humans , Imipenem/pharmacology , Klebsiella pneumoniae/drug effects , Meropenem , Microbial Sensitivity Tests , Multiplex Polymerase Chain Reaction , Phenotype , Thienamycins/pharmacology , Turkey , beta-Lactamases/genetics , beta-Lactams/pharmacology
5.
Microb Ecol ; 71(2): 339-46, 2016 Feb.
Article in English | MEDLINE | ID: mdl-26373644

ABSTRACT

The present study compared two chemical-based methods, namely, matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS) and attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy, to understand the misidentification of Exophiala dermatitidis and Exophiala phaeomuriformis. The study utilized 44 E. dermatitidis and 26 E. phaeomuriformis strains, which were partially treated with strong acids and bases for further evaluation. MALDI-TOF MS and ATR-FTIR spectroscopy data of the two Exophiala species were compared. Data groupings were observed for the chromic acid- and nitric acid-treated species when the black yeast sources were categorized as creosoted-oak sleepers, concrete sleepers, or dishwasher isolates. The MALDI-TOF MS data for the metalloenzyme-containing regions were consistent with the ATR-FTIR spectroscopy data. These results indicated that environmental isolates might contain metals not found in human isolates and might interfere with chemical-based identification methods. Therefore, MALDI-TOF MS reference libraries should be created for clinical strains and should exclude petroleum-associated environmental isolates.


Subject(s)
Exophiala/chemistry , Mycological Typing Techniques/methods , Phaeohyphomycosis/microbiology , Spectroscopy, Fourier Transform Infrared/methods , Tandem Mass Spectrometry/methods , Exophiala/classification , Exophiala/isolation & purification , Humans , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
6.
Med Mycol ; 53(4): 347-52, 2015 May.
Article in English | MEDLINE | ID: mdl-25851261

ABSTRACT

In this study, we investigated the applicability of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) for the identification of Exophiala species. The analysis included a total of 110 Exophiala isolates, including 15 CBS strains representing 4 species, Exophiala dermatitidis (61), E. phaeomuriformis (36), E. crusticola (9), and E. heteromorpha (4), that had been previously identified based on internal transcribed spacer (ITS) regions. We also compared the relative efficacies of Sabouraud glucose agar (SGA) and Columbia agar (CA) for use in MALDI-TOF MS. Remarkably, we obtained a log-score value ≥2.0 by using either SGA or CA for all 15 CBS strains, indicating species-level identification. The remaining 95 Exophiala strains were identified to the genus or species levels, with identification rates of 96.8% and 90.5%, using SGA or CA, respectively. Most of the E. dermatitidis (100% and 92.9%), E. phaeomuriformis (80.6% and 83.9%), E. crusticola (50% and 100%), and E. heteromorpha (100% and 100%) isolates were correctly identified using SGA or CA, respectively. Furthermore, 58.9% and 26.3% of the strains had log-score values of ≥2.0 by using SGA and CA, respectively. Our results indicate that MALDI-TOF MS is a rapid and reliable technique with high rates of correct taxonomic identification.


Subject(s)
Exophiala/chemistry , Exophiala/cytology , Microbiological Techniques/methods , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods , Culture Media/chemistry , Exophiala/growth & development , Time Factors
7.
Clin Lab ; 60(9): 1569-72, 2014.
Article in English | MEDLINE | ID: mdl-25291955

ABSTRACT

Organ transplant recipients under immunosuppressive therapy have a highly increased risk of opportunistic fungal infections. Cutaneous infection caused by Alternaria species are relatively rare in humans and most cases reported in the literature are in immunocompromised individuals. We report here on a 33-year old male renal transplant patient with diabetes mellitus who presented with cutaneous alternariosis caused by Alternaria infectoria, two years after the transplant. The diagnosis was performed by real-time polymerase chain reaction assay and histopathologic examination. The extension of the lesion under itraconazole treatment required treatment consisting of a combination of surgical excision and liposomal amphotericin B.


Subject(s)
Alternaria/genetics , Alternariosis/microbiology , Bacteriological Techniques , DNA, Fungal/isolation & purification , Kidney Transplantation/adverse effects , Opportunistic Infections/microbiology , Real-Time Polymerase Chain Reaction , Adult , Alternaria/classification , Alternaria/immunology , Alternaria/isolation & purification , Alternariosis/diagnosis , Alternariosis/immunology , Alternariosis/therapy , Humans , Immunocompromised Host , Immunosuppressive Agents/adverse effects , Male , Opportunistic Infections/diagnosis , Opportunistic Infections/immunology , Opportunistic Infections/therapy , Predictive Value of Tests , Time Factors , Treatment Outcome
8.
Mikrobiyol Bul ; 48(3): 429-37, 2014 Jul.
Article in Turkish | MEDLINE | ID: mdl-25052109

ABSTRACT

Hepatitis C virus (HCV) is one of the major causes of chronic hepatitis. It is important to know the genotypes of HCV in the decision of the HCV related chronic hepatitis therapy. The aim of this study was to evaluate the HCV genotypes determined at the Microbiology Laboratory of Akdeniz University Hospital, and to evaluate the changes in the distribution of the genotypes within the last five years. A total of 422 blood samples from HCV-RNA positive chronic hepatitis C patients (219 male, 203 female; age range: 8-79 yrs, mean age 46.3 ± 15.5 yrs) which were sent to our laboratory for genotyping between 2009-2013 period, were analyzed retrospectively. HCV-RNA extractions were performed in an automated system (EZ1 Virus Mini Kit v2.0, Qiagen, Germany), and a commercial reverse hybridization line probe-based assay (LIPA; GEN-C RT-PCR, Italy) was carried out for genotyping, For viral load determinations, a real-time PCR method (Cobas TaqMan HCV, Roche Diagnostics, Germany) was used. Demographic data of the patients were obtained from the hospital information systems and electronic patients' files. Out of the 422 patients, genotype 1b was detected in 63.3% (n= 267), genotype 1a in 14.7% (n= 62), genotype 3a in 11.1% (n= 47), genotype 2b in 0.9% (n= 4), genotype 4e in 0.2% (n= 1). The subtypes couldn't be determined for 5.4% (n= 23), 2.6% (n= 11) and 1.4% (n= 6) of the patients infected with genotype 1, 2 and 4, respectively. One (0.2%) patient, was coinfected with genotype 1 and 4. Of the patients, 40 were foreign-born (16 cases from Russia; 4 of each from Ukraine and Georgia; 3 of each from Turkmenistan, Kyrgyzstan, and Germany; one of each from Tajikistan, Azerbaijan, Uzbekistan, Chechnya, Moldova, Switzerland and Romania) and among these patients genotype 3a (19/40; 47.5%) was the most common genotype followed by genotype 1b (17/40; 42.5%). Median values of HCV viral load were 668.500 IU/ml (range: 2.000-9.630.000) in the whole group; while it was 732.000 IU/ml (range: 2.000-9.630.000) in patients infected with genotype 1 and 444.000 IU/ml (range: 2.650- 8.330.000) in patients infected with the other genotypes (p> 0.05). Patients infected with genotype 1 were found to be older than those infected with other genotypes (47 ± 15.7 and 39.5 ± 12.2, respectively; p< 0.001). Among patients infected with different genotypes, there was no statistically significant difference in terms of genders (p> 0.05). In conclusion, the determination of HCV genotypes is of crucial importance for treatment decision-making of chronic HCV infection. Besides, it also allows monitoring the changes in the epidemiology of HCV. In this study, although genotype 1b was determined as the most common HCV genotype, the detection of other genotypes was remarkable. This finding was attributed to the presence of many foreign national people in Antalya region which was a high capacity tourism area in Turkey.


Subject(s)
Genotype , Hepacivirus/classification , Hepatitis C, Chronic/virology , Adolescent , Adult , Age Distribution , Aged , Asia, Central/ethnology , Child , Europe/ethnology , Female , Hepacivirus/genetics , Hepatitis C, Chronic/epidemiology , Hepatitis C, Chronic/ethnology , Humans , Male , Middle Aged , Retrospective Studies , Russia/ethnology , Travel , Turkey/epidemiology , Young Adult
9.
Mikrobiyol Bul ; 48(1): 143-50, 2014 Jan.
Article in Turkish | MEDLINE | ID: mdl-24506725

ABSTRACT

Hepatitis B virus (HBV) and hepatitis C virus (HCV) infections are significant causes of morbidity and mortality in hemodialysis patients, since those patients are highly susceptible to infections due to immune suppression. The aims of this study were to investigate the presence of HBV and HCV infections in chronic hemodialysis patients by serological and molecular methods, and to determine the rate of occult HBV infection and the viral genotypes. A total of 201 patients who were under hemodialysis due to end-stage renal disease, were retrospectively evaluated. The study involved the patients at three different centers in Antalya, Turkey during 2006. HBV and HCV markers in the patients' sera were screened by ELISA method, viral nucleic acids were investigated by real-time polymerase chain reaction (PCR) in patients' plasma and viral genotypes were determined by DNA sequence analysis. Detection of at least one of the HBV markers HBsAg, anti-HBc total, and HBV DNA, was accepted as HBV infection, and detection of anti-HCV and/or HCV RNA was accepted as HCV infection. HBsAg positive patients with negative HBV DNA were considered as occult HBV infection. Of the patients 80 (40%) were female, 121 (60%) were male and the mean age was 51.16 ± 16.28 (range 17-93) years. In our study, sole anti-HBs positivity due to HBV vaccination, was detected in 89 (44.3%) patients. One hundred (50%) patients were found positive in terms of HBV infection and 40 (20%) were positive for HCV infection, while 24 (12%) patients had HBV and HCV co-infections. Eighty-five (42.3%) patients had no HBV and HCV infection. Among the 5 (2.5%) patients who were HBsAg positive, four were also HBV DNA positive. Occult HBV infection was detected in 1 (0.5%) patient. Anti-HCV and HCV RNA were found positive in 37 (18.4%) and in 24 (12%) patients, respectively. Among the HCV-RNA positive patients, 3 (12.5%) were anti-HCV negative. ALT and AST levels were found normal in all of the HBV DNA positive patients, and 62.5% (15/24) of HCV RNA positive patients. All of the HBV isolates were identified as genotype D and HCV isolates as genotype 1b. No statistically significant correlation was detected between the HBV infection and patients' age, duration of hemodialysis and elevation of serum transaminase levels (p> 0.05). On the other hand, HCV infection was seen to increase with age (p= 0.047). HCV infection showed a statistically significant increase with the duration of hemodialysis. HCV infection risk was increased in patients who were under hemodialysis for ≥ 25 months (p< 0.001, OR: 0224, 95% CI= 0089-0562). There was also a statistically significant correlation between the presence of HCV infection (anti-HCV and/or HCV RNA positive) and high levels of serum transaminases (p< 0.001). However, in two of the three cases who were anti-HCV negative and HCV RNA positive, serum transaminase levels were normal while the viral loads were high. Therefore to follow-up HCV infection in the hemodialysis patients, anti-HCV and serum transaminase levels may not be sufficient alone and these patients should be evaluated periodically for HCV RNA. In addition, the detection of occult HBV infection in one of the study patients, indicated that HBV DNA should also be investigated at regular intervals in the hemodialysis patients.


Subject(s)
Hepacivirus/isolation & purification , Hepatitis B virus/isolation & purification , Hepatitis B/diagnosis , Hepatitis C/diagnosis , Renal Dialysis/adverse effects , Adolescent , Adult , Age Factors , Aged , Aged, 80 and over , Enzyme-Linked Immunosorbent Assay , Female , Genotype , Hepacivirus/classification , Hepacivirus/genetics , Hepacivirus/immunology , Hepatitis B/etiology , Hepatitis B virus/classification , Hepatitis B virus/genetics , Hepatitis B virus/immunology , Hepatitis C/etiology , Humans , Immunocompromised Host , Kidney Failure, Chronic/therapy , Male , Middle Aged , Real-Time Polymerase Chain Reaction , Retrospective Studies , Sequence Analysis, DNA , Young Adult
10.
Turk J Med Sci ; 44(3): 476-83, 2014.
Article in English | MEDLINE | ID: mdl-25558652

ABSTRACT

AIM: Zygomycosis is a severe angioinvasive infection caused by Zygomycetes. We retrospectively investigated 16 cases of zygomycosis. MATERIALS AND METHODS: The data of patients, who had been followed between 2004 and 2010 in 8 tertiary-care teaching hospitals, were reviewed. Demographic characteristics, underlying diseases, and clinical signs and symptoms of the patients, as well as diagnostic methods, data obtained by radiological imaging methods, and the therapies, were recorded. Therapeutic approaches, antifungal agents and duration of use, and the characteristics of the cases were identified. RESULTS: The study included 11 female and 5 male subjects. The most common symptoms and clinical signs were fever (n = 9) and retro- orbital pain (n = 7). Rhinocerebral zygomycosis was the most common form. The mean time elapsed for diagnosis was 14.26 + 13.96 (range: 2-52) days. Antifungal therapy was given to 15 patients (94%). In addition to antifungal therapy, 12 patients underwent surgical intervention 1 to 4 times. The mean duration of receiving antifungal therapy was 61.4 + 58.02 (range: 1-180) days. The median duration of treatment was 62.5 (range: 42-180) days in survivors. CONCLUSION: Zygomycosis is an infectious disease with high mortality despite antifungal therapy and surgical interventions.


Subject(s)
Zygomycosis/diagnosis , Adult , Aged , Antifungal Agents/therapeutic use , Female , Humans , Male , Middle Aged , Retrospective Studies , Young Adult , Zygomycosis/drug therapy
11.
Ann Lab Med ; 33(5): 326-30, 2013 Sep.
Article in English | MEDLINE | ID: mdl-24003422

ABSTRACT

BACKGROUND: Active screening for vancomycin-resistant enterococci (VRE) using rectal specimens is recommended to limit the spread of antimicrobial resistance within certain high-risk populations. We evaluated the diagnostic performance of Vancomycin Resistance 3 Multiplexed Tandem PCR assay (AusDiagnostics, Australia), a rapid multiplex real-time PCR assay that detects vanA and/or vanB. METHODS: Two-hundred-and-eleven rectal swabs from Hematology and Oncology unit were submitted for VRE surveillance via direct detection of vanA and/or vanB by culture and by using Vancomycin Resistance 3 Multiplexed Tandem PCR assay. Enterococci were identified to the species level by using standard biochemical tests and BD Phoenix Automated Microbiology System (BD Diagnostic Systems, USA). Vancomycin susceptibility of enterococci was determined using Etest (BioMerieux, France). RESULTS: Compared to the culture method, Vancomycin Resistance 3 Multiplexed Tandem PCR assay had a sensitivity of 84.0%, specificity of 98.8%, positive predictive value (PPV) of 91.3%, and negative predictive value (NPV) of 97.6%. The assay failed to detect 18 (8.5%) specimens because of the presence of PCR inhibitors; of the remaining 193 specimens, 25 (12.9%) were positive, 23 for vanA, and 2 for vanB. Although both sensitivity and specificity for vanA VRE was 100% compared to the culture method, all vanB-positive specimens tested negative by VRE culture. CONCLUSIONS: Vancomycin Resistance 3 Multiplexed Tandem PCR assay is a rapid and laborsaving option for VRE surveillance for direct use on rectal swabs. However, the high rate of PCR failure owing to the inhibitors in the specimens and the low specificity for vanB should be considered when interpreting the results.


Subject(s)
DNA, Bacterial/analysis , Enterococcus/drug effects , Enterococcus/genetics , Multiplex Polymerase Chain Reaction , Rectum/microbiology , Vancomycin Resistance/genetics , Vancomycin/pharmacology , Bacterial Proteins/genetics , Carbon-Oxygen Ligases/genetics , Enterococcus/growth & development , Enterococcus/metabolism , Humans , Reagent Kits, Diagnostic , Sensitivity and Specificity
12.
Mikrobiyol Bul ; 47(1): 141-6, 2013 Jan.
Article in Turkish | MEDLINE | ID: mdl-23390911

ABSTRACT

Enterococci which are part of the commensal flora of the human gastrointestinal and genitourinary tracts, are increasing in importance as the cause of hospital-acquired infections. Identification of Enterococcus spp. at the species level is of great importance, for appropriate treatment of patients, infection control and to supply epidemiological data. Conventional methods for the identification of enterococcus isolates at species level is difficult and time consuming. Correct identification of enterococcus isolates in clinical microbiology laboratory by conventional methods is replaced by semi-automated or automated identification and molecular methods. The aim of this study was to evaluate the performance of Phoenix automated system (BD Diagnostic Systems, USA), API Rapid ID 32 Strep System (bioMerieux, France) and Enterococcus MGRADE LightCycler kit (Roche Molecular Biochemicals, Germany) used in real-time polymerase chain reaction (Rt-PCR), for the species level identification of enterococcus strains isolated from clinical specimens. A total of 90 vancomycin susceptible enterococci isolated from different patients were identified by all of the three commercial systems, together with conventional methods. Of the strains, 59 were identified as E.faecalis, 28 were E.faecium, and one of each as E.raffinosus, E.hirae and E.casseliflavus with conventional methods. One E.faecalis strain identified by the conventional system was identified as E.faecium by Phoenix system and one E.faecium strain as E.durans. One E.raffinosus strain identifed by the conventional method was identified as E.avium by API. Conventionally identified four E.faecalis strains were determined to be E.faecium by Rt-PCR and one E.faecium, one E.raffinosus and one E.casseliflavus as E.faecalis. Accordingly, the consistency of Phoenix, API Rapid ID 32 Strep and LightCycler Enterococcus MGRADE systems with the conventional methods were detected as 97.8% (88/90), 98.9% (89/90), and 92.2% (83/90), respectively. In conclusion, all of those three commercial assays are appropriate methods to be used for the identification of enterococci at the species level in the routine clinical microbiology laboratories, due to their high compliance with the conventional method, and their ability to yield the results at the same day.


Subject(s)
Enterococcus , Vancomycin , Cross Infection/microbiology , Enterococcus/isolation & purification , Germany , Gram-Positive Bacterial Infections/microbiology , Humans , Microbial Sensitivity Tests , Polymerase Chain Reaction , Streptococcal Infections
13.
Med Mycol ; 50(5): 529-32, 2012 Jul.
Article in English | MEDLINE | ID: mdl-21988703

ABSTRACT

The aim of the study was to investigate the distribution of Candida species isolated from urine specimens of hospitalized patients in Akdeniz University Hospital, Antalya, Turkey, as well as their susceptibilities to antifungal agents. A total of 100 patients who had nosocomial candiduria between March 2003 and May 2004 at the facility were included in the study. Organisms were identified by conventional methods and the use of API ID 32C strips. Susceptibilities of the isolates to amphotericin B were determined by Etest, whereas the minimum inhibitory concentration (MIC) values of these same strains to fluconazole, voriconazole and caspofungin were assessed using the broth microdilution method. The most common species recovered was C. albicans 44% of all yeasts, followed by C. tropicalis (20%), C. glabrata (18%), C. krusei (6%), C. famata (5%), C. parapsilosis (4%), C. kefyr (2%) and C. guilliermondii (1%). A total of nine (9%) of the isolates, including five C. krusei and four C. glabrata isolates were susceptible dose-dependent (SDD) to fluconazole. In constrast, only two C. glabrata and one C. krusei isolates were resistant to this antifungal. The voriconazole MICs for all Candida isolates were ≤0.5 µg/ml, except for one C. glabrata isolate with a MIC value of 2 µg/ml. Among all isolates, 94% were susceptible to amphotericin B with MIC values of <1 µg/ml and all isolates were susceptible to caspofungin with MIC values of ≤0.5 µg/ml. Future studies are needed to define better treatment regimens for those patients who have fluconazole-resistant Candida urinary tract infections.


Subject(s)
Antifungal Agents/pharmacology , Candida/drug effects , Candida/isolation & purification , Candidiasis/microbiology , Cross Infection/microbiology , Urinary Tract Infections/microbiology , Adolescent , Adult , Aged , Aged, 80 and over , Candida/classification , Candidiasis/drug therapy , Child , Child, Preschool , Cross Infection/drug therapy , Drug Resistance, Fungal , Female , Fluconazole/pharmacology , Fluconazole/therapeutic use , Hospitals, University , Humans , Infant , Infant, Newborn , Male , Microbial Sensitivity Tests , Middle Aged , Species Specificity , Turkey , Urinary Tract Infections/drug therapy , Urine/microbiology , Young Adult
14.
Mikrobiyol Bul ; 44(3): 367-74, 2010 Jul.
Article in Turkish | MEDLINE | ID: mdl-21063986

ABSTRACT

The aim of this study was to determine the extended-spectrum beta-lactamase (ESBL) types by isoelectric focusing (IEF) and polymerase chain reaction (PCR) methods in 56 Escherichia coli strains isolated from urine samples of patients with community-acquired urinary tract infection and determined as ESBL positive with the phenotypic screening tests (E test and combined disk method). IEF revealed that most of the strains produced 1 to 3 different bands, mostly at the isoelectric points 8.2 (n= 44, 79%) compatible with CTX-M. Twenty four (43%) isolates had CTX-M and TEM enzyme bands together, 16 (29%) isolates had only CTX-M enzyme bands, 3 (5%) isolates had CTX-M, TEM, SHV bands, one had CTX-M and SHV enzyme bands together, and one had only TEM band. Eleven E.coli strains did not yield any enzyme bands. PCR analysis revealed that 93% (n= 52) of the isolates had CTX-M, 64% (n= 36) had TEM and 11% (n= 6) had SHV, while 29 (52%) had CTX-M + TEM, three had CTX-M + SHV, and three had CTX-M + TEM + SHV genes together. PER-1 type beta-lactamases were not detected by PCR method. PCR analysis of the eleven strains that yielded no band in IEF showed that 5 strains had CTX-M + TEM, 3 had CTX-M and 3 had TEM enzyme genes. The consistency between IEF and PCR methods for the determination of CTX-M, TEM and SHV enzymes was 85%, 78% and 67%, respectively. Genes encoding ESBL's are usually located on transferrable plasmids that may also carry other resistance determinants. Thus detection of beta-lactamase enzyme types in ESBL positive bacteria is important for the choice of appropriate antimicrobial agents for treatment.


Subject(s)
Escherichia coli Infections/microbiology , Escherichia coli/enzymology , Urinary Tract Infections/microbiology , beta-Lactamases/metabolism , Community-Acquired Infections/microbiology , Escherichia coli/classification , Escherichia coli/drug effects , Escherichia coli/genetics , Humans , Isoelectric Focusing , Polymerase Chain Reaction , beta-Lactamases/chemistry , beta-Lactamases/genetics
15.
Pol J Microbiol ; 59(1): 55-60, 2010.
Article in English | MEDLINE | ID: mdl-20568531

ABSTRACT

Brucellosis is a zoonosis with a worldwide distribution and remains a significant public health problem mainly in the developing world. In this study we evaluated the in vitro activities and synergistic effects of antibiotic combinations against blood culture isolates of Brucella spp. In vitro susceptibilities of 76 blood culture isolates of Brucella melitensis and one blood culture isolate of Brucella abortus to doxycycline, streptomycin, gentamicin, trimethoprim-sulfamethoxazole, moxifloxacin, rifampin, ciprofloxacin, and tigecycline were examined by Etest method. For 37 patients with Brucella spp. isolates (36 B. melitensis, 1 B. abortus), antibiotic combinations used for treatment were identified with those tested in vitro for synergy using Etest method. Trimethoprim-sulfamethoxazole and tigecycline were the most active of the compounds tested with MIC90 value of 0.094 mg/l. Among antibiotic combinations only streptomycin-rifampin combination was synergistic for one Brucella spp. isolate. The other antibiotic combinations revealed antagonistic or indifferent activity. Complete clinical response was achieved in all patients. Further studies are required to determine the correlation between the antimicrobial susceptibility and synergy test results with the clinical course of patients. Brucellosis can be adequately treated with existing regimens in our region.


Subject(s)
Anti-Bacterial Agents/pharmacology , Brucella/drug effects , Minocycline/analogs & derivatives , Adolescent , Adult , Aged , Brucellosis/drug therapy , Child, Preschool , Drug Synergism , Female , Humans , Male , Microbial Sensitivity Tests , Middle Aged , Minocycline/pharmacology , Tigecycline
17.
J Clin Microbiol ; 47(1): 278-81, 2009 Jan.
Article in English | MEDLINE | ID: mdl-19005150

ABSTRACT

Fusarium species are saprophytic molds which cause disseminated or localized infections in humans. Disseminated Fusarium infection can cause significant morbidity and mortality in immunocompromised patients. We present a case of disseminated fusariosis caused by Fusarium verticillioides in a patient with acute lymphoblastic leukemia and successfully treated using both liposomal amphotericin B and voriconazole.


Subject(s)
Fusarium/isolation & purification , Hematopoietic Stem Cell Transplantation/adverse effects , Mycoses/diagnosis , Precursor Cell Lymphoblastic Leukemia-Lymphoma/complications , Amphotericin B/therapeutic use , Antifungal Agents/therapeutic use , Child , Humans , Immunocompromised Host , Male , Mycoses/drug therapy , Mycoses/microbiology , Precursor Cell Lymphoblastic Leukemia-Lymphoma/therapy , Pyrimidines/therapeutic use , Triazoles/therapeutic use , Voriconazole
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