[Investigation of the VISA and hVISA Presence in Methicillin-Resistant Staphylococcus aureus Isolates by Various Methods]. / Metisiline Dirençli Staphylococcus aureus Izolatlarinda Çesitli Yöntemlerle VISA ve hVISA Varliginin Arastirilmasi.

Staphylococcus aureus is an important human pathogen that causes community and hospital-acquired infections. The role of vancomycin in the treatment of methicillin-resistant S.aureus infections is indisputable. However, vancomycin intermediate susceptible S.aureus (VISA) and heterogeneously VISA (hVISA) isolates, that cause treatment failures during the use of vancomycin, cannot be detected by routine laboratory methods. The gold standard method for the detection of these isolates is the population profile analysis-area under the curve (PAP-AUC) method. In this study, it was aimed to determine the presence of mecA and mecC gene regions that cause methicillin resistance, the clonal relationship between isolates, and the presence of VISA and hVISA. A total 68 methicillin-resistant S.aureus (MRSA) strains were included in this study which were isolated in the microbiology laboratory of the hospital between 2015- 2020. Identification of the isolates were determined by matrix assisted laser desorption ionization-time of flight mass spectrophotometry (VITEK MS, BioMérieux, France). Methicillin resistance was investigated by disk diffusion method using cefoxitin (30 µg, Bioanalyse, Türkiye) disk and vancomycin MIC values were determined by broth microdilution method. mecA and mecC gene regions were investigated by polymerase chain reaction (PCR) method. The presence of VISA and hVISA were investigated by modified agar screening, macro gradient diffusion test and confirmated by PAP-AUC methods, and the clonal relationship between isolates were investigated by pulsed field gel electrophoresis method. The mecA gene region was determined in all isolates, but the mecC gene region was not found in any of the isolates. The MIC50 value of the isolates was determined as 1 µg/mL and the MIC90 value was determined as 2 µg/mL by broth microdilution method. Six VISA and four hVISA suspected strains were detected by a modified agar screening method. Among the isolates identified as suspicious by the modified agar screening method, one isolate was evaluated as VISA and one isolate was evaluated as hVISA by the gold standard PAP-AUC method. No dominant epidemic isolate has been identified by PFGE. As a result, VISA and hVISA were determined in the hospital. The increase in these isolates is a serious concern. For this reason, it is believed that it would be beneficial to investigate the VISA/hVISA ratios in MRSA isolates at certain periods, and to take necessary infection control measures to implement measures and practices to prevent the spread of these isolates in the community and hospital environment.

[Co-production of OXA-48 and NDM-1 Carbapenemases in Providencia rettgeri: the first report]. / Providencia rettgeri'de OXA-48 ve NDM-1 karbapenemaz genlerinin birlikte üretimi: ilk bildirim.

Our country is the epicenter of the OXA-48-like carbapenemase-producing Klebsiella and Escherichia coli; and in the recent years, the concern has been increasing due to both spreading of this resistance to other members of Enterobacteriaceae family and acquiring other carbapenemases by the OXA-48-producing strains. In this study, OXA-48 and NDM-1 co-production was presented in Providencia rettgeri. Two P.rettgeri strains that were resistant to all antimicrobials except colistin and tigecyclin, were isolated from two patients in the burn unit of our hospital, including one from the urine sample of a 68 years female in April 2017, and the other from a burn wound swab of a 35 years old male, in November 2017. Minimal inhibitory concentrations (MICs) of the isolates for imipenem and meropenem were measured as ≥ 32 µg/ml; and for colistin and tigecyclin were 1 ve 0.5 µg/ml, respectively. Multiplex PCR analysis showed that both strains were carrying blaOXA-48 and blaNDM-1 carbapenemases, and blaTEM extended spectrum beta-lactamase genes. By using DNA sequence analysis, the TEM gene was typed as blaTEM-1. The Pulsed Field Gel Electrophoresis (PFGE) analysis indicated that these two strains which were consecutively isolated from two different patients in a single unit within about seven months were genetically indistinguishable. No significant data that could explain the spread of these isolates was obtained from our retrospective analysis of the medical records including the results of environmental surveillance cultures, and patients' history. Nevertheless, hospital infection control committee enforced the infection control measures in that unit, and no further isolation was observed within three months period following the last isolation, neither from environmental nor from clinical samples. With this study, it was emphasized that the co-production of OXA-48 and NDM-1 carbapenemases which was reported from only three Enterobacteriaceae species up to date was ongoing for spreading to other species by using horizontal route, and also showing a potential to be a growing problem in the hospitals, by clonal expansion (vertical route). Effectively using of the molecular epidemiological methods will provide useful data to better understand the transmission dynamics of such rare, but problematic species in hospitals.

[Evaluation of the diagnostic performance of Xpert MTB/RIF test for the detection of Mycobacterium tuberculosis and rifampin resistance in clinical samples]. / Klinik örneklerde Mycobacterium tuberculosis'in saptanmasi ve rifampin direncinin tespitinde Xpert MTB/RIF testinin tanisal performansinin degerlendirilmesi.

Rapid and accurate detection of active tuberculosis (TB) cases is one of the most important goal of tuberculosis control programme. For this purpose, new methods are being developed to isolate, serotype and determine the drug resistance of the agent. Xpert MTB/RIF test (CepheidGeneXpert® System, USA) that has been recently developed, is a real-time polymerase chain reaction-based method which detects Mycobacterium tuberculosis complex and resistance of the strain to rifampicin (RIF) from the clinical sample directly within a couple of hours. However, there are not sufficient data about the performance of that test for extrapulmonary samples and pulmonary samples other than sputum. The aims of this study were to investigate the sensitivity, specificity, positive and negative predictive values of Xpert MTB/RIF test in detection of M. tuberculosis and the performance in the determination of rifampicin resistance of the isolates from pulmonary and extrapulmonary clinical samples. A total of 2160 clinical samples, in which 1141 (52.8%) were pulmonary and 1019 (47.2%) were extrapulmonary samples, sent to our laboratory between July 2013 to December 2014, were included in the study. Sixty seven of the evaluated samples (3.1%) were positive with microscopy (acid-fast stain; AFS), 116 samples (5.1%) were positive with culture and 98 samples (4.5%) were positive with Xpert MTB/RIF test. When the culture was considered as the reference method, the sensitivity and specificity of Xpert MTB/RIF test were determined as 73.3% and 99.3%, respectively for all samples; 77.5% and 99.5%, respectively for pulmonary samples and 63.9% and 99.2%, respectively for extrapulmonary samples. Among AFS positive samples, the sensitivity was 100% and specificity was 66.7%; whereas among AFS negative samples those values were 40.4% and 99.4%, respectively. Among all the samples involved in the study, RIF resistance was determined only in three samples with Xpert MTB/ RIF test and that was also proved phenotypically (100% concordance). According to mycobacterial culture results, positive and negative predictive values of Xpert MTB/RIF test were determined as 86.7% and 98.5%, respectively for all samples. Those were determined as 92.5% and 98.3%, respectively for extrapulmonary samples and were determined as 74.2 and 98.7%, respectively for pulmonary samples. According to the results obtained in our study, sensitivity of Xpert MTB/RIF test for extrapulmonary samples was found to be at moderate level; sensitivity of the test was found to be decreased especially in AFS negative samples with less bacilli load. Nonetheless, specificity of Xpert MTB/RIF test to the agent in all samples was found to be extremely high. In our study, although RIF-resistant strains were detected in few of the samples, Xpert MTB/ RIF test could differentiate all resistant and sensitive strains. Additionally, detection of M. tuberculosis and RIF resistance in our laboratory takes approximately 20.96 days with culture, this period decreases to a couple of hours with Xpert MTB/RIF test. Because of the advantages such as being practical, rapid and requiring minimal safety measures, it was concluded that Xpert MTB/RIF test may contribute to rapid diagnosis of TB also in extrapulmonary samples, with the confirmation of culture method.

Oral colonization of Acinetobacter baumannii in intensive care units: Risk factors, incidence, molecular epidemiology, association with the occur of pneumonia and sepsis, and infection control measures.

Objectives: Oral colonization of Acinetobacter baumannii can lead to infections such as pneumonia and sepsis. We aimed to evaluate oral colonization of hospitalized patients in ICUs and to examine risk factors for oral colonization, molecular epidemiology, and incidence of pneumonia and sepsis. Materials and Methods: The study began in February 2021. Oral cultures were taken. The microorganisms were identified by a Maldi-tof MS mass spectrometry device. Colistin resistance genes were investigated by polymerase chain reaction. Clonal relationships were determined by pulsed-field gel electrophoresis. Results: A. baumannii was found in 21 of 96 patients' oral cultures. Pneumonia and sepsis due to A. baumannii were detected in 14 and 5 patients, respectively. The mean growth time of A. baumannii from oral cultures was 11.8 days, and the meantime for the occurrence of pneumonia after oral growth was 5.2 days. We determined a plasmid mediated mcr-2 colistin resistance gene in a colistin susceptible A. baumannii strain. It is the first report of the plasmid mediated mcr-2 colistin resistance gene in our country. In total, fourteen different A. baumannii genotypes were determined in PFGE. It was determined that the effects of antibiotic use, oral motor dysfunction, mechanical ventilation, intubation, orogastric tube use, and total parenteral nutrition intake on oral colonization were statistically significant. Conclusion: Oral colonization of A. baumannii is a significant concern in ICUs. We believe that it is important to take oral cultures and follow the risk factors and take infection control measures to prevent oral colonization of resistant isolates in ICUs.

[Investigation of hydrophobic characteristics of biofilm producer and non-producer Staphylococcus aureus clinical isolates]. / Biyofilm olusturan ve olusturmayan Staphylococcus aureus klinik izolatlarinin hidrofobik ozelliklerinin arastirilmasi.

The ability of staphylococcus to adhere certain structures and to form biofilm (slime) layer plays an important role in the pathogenesis of staphylococcal infections. Hydrophobic interactions and hydrogen bonds are important factors that play role in adherence. This study was designed to compare the hydrophobic properties of slime positive and negative Staphylococcus aureus strains isolated from blood cultures. Ten methicillin-resistant S. aureus isolates (five of them being slime positive) obtained from blood cultures of patients at intensive care unit of a university hospital, between May 2006 and June 2007, were included in the study. Slime production of the isolates was determined by Christensen's method. Methicillin resistance was determined by cefoxitin disc test and oxacillin salt agar test. It was determined that the test strains did not exhibit any autoaggregation. The adherence of strains to the three different hydrocarbons as solid phases (butyl-sepharose, octyl-sepharose and phenyl-sepharose; Amersham Bioscience, Sweden) were studied by using hydrophobic interaction chromatography (HIC) method. After butyl- and octyl-sepharose chromatography, it was determined that slime negative S. aureus strains were separated into three fractions eluted with phosphate buffered saline (PBS), 40% and 96% ethanol, while slime positive strains were separated into two fractions eluted with 40% and 96% ethanol, respectively. By phenyl-sepharose chromatography analysis; both slime negative and positive strains were separated into two fractions eluted in 40% and 96% ethanol. Hydrophobicity tests were repeated at 4 degrees C and pH 6-9 to evaluate the effect of changing conditions on hydrophobicity. However, no changes we re observed at these temperature and pH values. According to these analysis it was concluded that; (a) S. aureus strains consist heterogeneous fractions with distinct hydrophobic binding strengths; (b) hydrophobic surface protein secretion may be different in heterogeneous groups, and (c) slime positive S. aureus strains were more hydrophobic than non-slime producing strains. Further research is required in order to characterise the eluted fractions and to evaluate their pathogenic capacities.

Bacterial translocation to kidney in rats with intestinal obstruction and the role of nitric oxide.

OBJECTIVE: Bacterial translocation (BT) is the passage of viable indigenous bacteria from one site to another, such as from gastrointestinal tract to the normally sterile regional mesenteric lymph nodes and than other internal organs. In this study we aimed to investigate the BT to kidney and the protective effect of nitric oxide (NO) inhibition. MATERIAL AND METHODS: A total of 40 adult male Wistar albino rats weighing 320-350g were divided into four equal groups. Group 1 (n = 10): control group, group-2 (n = 10) sham control, group-3 (n = 10) simple obstruction, in which ileum was ligated 1-2 cm proximal to the ileocecal valve, group-4 (n = 10), simple obstruction and treated with L-NAME. Twenty four hour after the operation rats were sacrificed and kidneys were removed by sterile manner and trunk blood obtained for NO analysis. BT was defined as any positive culture from the blood and kidney. Results were compared with Mann- Whitney U test. RESULTS: NO levels in control, sham group, simple obstruction group and obstruction plus L-NAME treated group were 14.04 +/- 0.65 micromol/L, 13.03 +/- 0.080 micromol/L, 31.17 +/- 0.40 micromol/L and 12.24 +/- 0.70 micromol/L, respectively. Renal culture results were negative in all controls and sham operated rats. However, all culture results were positive in obstruction group and in 4 in L-NAME-treated group. The most common microorganism that translocated was E. coli. CONCLUSION: This experimental study shows that BT to rat rat kidneys occurs in bowel obstruction and this can be inhibited by a NO inhibitor, L-NAME. Further studies are needed to define the clinical significance of these findings on urinary tract infection.

Molecular epidemiology of methicillin-resistant Staphylococcus aureus isolates from clinical specimens of patients with nosocomial infection: are there unnoticed silent outbreaks?

Bacteriological and epidemiological studies were carried out on 90 isolates of methicillin-resistant Staphylococcus aureus (MRSA) at Turgut Ozal Medical Center of Inönü University, (Malatya/Turkey). MRSA isolates were obtained from patients with nosocomial infections. Staphylococcus aureus clinical isolates were collected between May 2004-May 2005. Isolates were tested for resistance to methicillin. Antimicrobial susceptibility testing and slime production evaluation was performed. Genotype studies were carried out by arbitrarily primed polymerase chain reaction (AP-PCR) and consequent cluster analysis. All of the isolates were mecA-positive in a PCR-based assay; all exhibited resistance to oxacillin, by agar dilution (MICs > or = 4 mg/L) and disc diffusion methods, and multiple antibiotics. Most MRSA isolates were collected in intensive care units. Of 90 samples, 53 were found to be unrelated to the others while the remaining 37 strains were either identical or closely related. Dendrogram analysis identified nine major clusters. These data support the opinion that MRSA are significant nosocomial pathogens in intensive care units and that resistant clones may be transmitted between patients. Molecular epidemiological tools are helpful for understanding transmission patterns and sources of infection, and are useful for measuring outcomes of intervention strategies implemented to reduce nosocomial MRSA.

Laboratory-acquired skin infections in a clinical microbiologist: Is wearing only gloves really safe?

Laboratory-acquired infection is one of the leading occupational health hazards. On a laboratory worker's hands, carbuncles occurred. Staphylococcus aureus was isolated from pus samples of the carbuncles, with the same pulsed field gel electrophoresis band pattern with one of the recently studied strains in the laboratory. Incorrect or inadequate application of infection control measures may result in pathogen acquisition from the clinical samples, and wearing only gloves is not sufficient for the biosafety of laboratory workers in clinical diagnostic laboratories.

Association of Helicobacter pylori infection with systemic inflammation in preeclampsia.

OBJECTIVE: The aim of this study was to compare C-reactive protein (CRP), tumor necrosis factor alpha (TNFalpha), Chlamydia pneumonia IgG, IgM and plasma Helicobacter pylori IgA levels between preeclamptic and normal pregnant women and to determine whether seropositivity to Helicobacter pylori is associated with elevated levels of CRP and TNF-alpha. METHODS: Forty patients with preeclampsia and 40 normotensive pregnant women of similar age and body mass index at the third trimester of gestation were selected for the study. Chlamydia pneumonia IgM and IgGs, Helicobacter pylori IgAs and concentrations of CRP and TNF-alpha were measured. RESULTS: Concentrations of CRP and TNF-alpha were significantly higher in patients with preeclampsia than in control subjects. In the preeclamptic group, positivity rate for Helicobacter pylori IgA was significantly higher as compared to controls (p = 0.034). CRP and TNF-alpha levels were higher in Helicobacter pylori seropositive subjects. CONCLUSION: We demonstrated high levels of serum CRP and TNF-alpha in preeclamptic women who were seropositive to Helicobacter pylori in comparison with those in seronegative subjects.