The Evaluation of the Staphylococcus aureus Strains by Using Arbitrarily Primed-Polymerase Chain Reaction and Plasmid Profile Analysis Techniques

A total of 30 methicillin resistant Staphylococcus aureus (MRSA) strains were isolated as nosocomial pathogens from patients hospitalized in General Surgery, Orthopedics and Reconstructive Surgery Departments from June 1997 through December 1997. These strains were evaluated by moleculer typing methods using arbitrarily primed-polymerase chain reaction (AP-PCR) and plasmid profile analysis (PPA). By using AP-PCR method, these isolates on the basis of their band patterns were divided into seven groups (I through VII) in which each group had reciprocal number of isolates by thirteen, ten, two, two, one, one and one, respectively. By PPA, these isolates were grouped into three divisions as group A, B and C and each was represented by twentytwo, three and five isolates respectively. It was found that as nosocomial pathogens group I and II (77%) are frequently encountered for AP-PCR, while group A for PPA (73%) dominated

Cloning of Hepatitis B Virus Surface Gene Region to Escherichia coli

In this study, we cloned HBV s gene to E. coli by using PCR-dependent TA cloning procedure. Our aim was to achieve gene transfer via general principles of recombinant DNA technologies and to use the cloned DNA fragments in future expression studies to obtain antigens to be utilized for diagnostic and immunization procedures

Microorganisms and Antibiotic Resistance Patterns of Pneumonia in Patients on Mechanical Ventilation

To determine the causative microorganisms, their antibiotic resistance and empirical treatment in ventilator associated pneumonia (VAP), 101 patients on mechanical ventilation were studied. The material extracted by bronchoscopic techniques (protected specimen brush, bronchoalveolar lavage) was tested by classic microbiological and quantitative culture methods. VAP was detected at 45 (44.5%) of our patients. We isolated P. aeruginosa (35%), S. aureus (22.8%) and K. pneumoniae (17.5%) as the causative agents of VAP. P. aeruginosa isolates were found to be sensitive to ciprofloxacine (82%), amikacin (70%) imipenem (64%) and ceftazidime (60%), but were resistant to cefoperazone (88%), mezlocillin (85%) and gentamicin (80%) by standart disk diffusion method according to National Committee for Clinical Laboratory Standards (NCCLS) criteria. Staphylococcus aureus isolates were sensitive to cotrimoxazole (65%), ciprofloxacine (63%); methicillin resistance were found as 26%, all isolates were sensitive to vancomycin and teicoplanin. K. pneumoniae isolates were sensitive to ciprofloxacine (83%), amikacin (79%), imipenem (74%), ceftazidime (70%), but were resistant to ampicillin/sulbactam and cefazoline (80%), and tobramycin (69%). In conclusion, the causative microorganisms of VAP and their resistance patterns for each hospital should be taken into consideration for planning the empirical therapy

Antibody Seroprevalence of Diphtheria in Young Adults

Our study has been planned to search for the susceptibility of the young adult population to diphtheria. For this purpose, 410 soldiers in 20-25 age group have been tested for their antibodies against diphteriae. We have also recorded their ages, education status and geographic regions. Blood samples were taken from a research group of mean age 20.89 ± 0.095 and then 0.5 mL adult type diphtheria-tetanus vaccine (Imogam dT, Pasteur-Merieux, France) was applied. Of 410, the second blood samples of 316 subjects were taken for searching the diphtheria antibodies. Diphtheriae antibody levels were tested by using of Gen-enzyme Virotech GmbH (Löwenplatz 5, Rüsselsheim, Germany) test kit in the first and the second blood samples of these 316 subjects. 24.5% (50/204) of the 20 years age group, 29.4% (30/102) of the 21-24 years age group and 30% (3/10) of the 25 years age group had antibodies under the protective level of 0.01 IU/mL. Prior to vaccination period a statistically insignificant difference between the groups was determined with respect to age, educational status and geographic area (p> 0.05 for all groups). After the vaccination, in all age groups the antibody levels were found as above the protective level. According to these results, it can be concluded that dT vaccination should be considered for the 20 years age group

Investigation of extensive drug resistance in multidrug resistance tuberculosis isolates

Son yıllarda tüm dünyada ilaca dirençli tüberküloz (TB) olgularında izlenen artış, halk sağlığını önemli derecede tehdit etmektedir. Yaygın ilaca dirençli TB (YİD-TB), birinci seçenek anti-TB ilaçlardan izoniazid ve rifampisin direncine [çok ilaca dirençli TB (ÇİD-TB) tanımı] ek olarak herhangi bir florokinolona ve parenteral verilen ikinci seçenek anti-TB ilaçlardan en azından birisine direnç gelişmesi durumudur. Birinci seçenek anti-TB ilaçlardan olumlu yanıt alınamayan ÇİD-TB olgularında uygulanan anti-TB tedavi, klinik izolatların varsa ikinci seçenek anti-TB ilaç in vitro duyarlılık test sonuçlarına, yoksa standart tedavi protokollerine göre yapılmaktadır. Bireysel anti-TB tedavinin, ideal olarak izolatın duyarlılık test sonuçlarına bakılarak yapılması önerilse de, ikinci seçenek anti-TB ilaç duyarlılık testlerinin sıklıkla yapılamadığı ülkemiz gibi gelişmekte olan ülkelerde her zaman standart tedavi protokollerine gereksinim duyulmaktadır. Bu nedenle, uygun standart tedavi protokollerinin oluşturulabilmesi için ulusal ve bölgesel ölçekli direnç paternlerini ortaya koyacak çalışmalara ihtiyaç bulunmaktadır. Bu çalışmada, İstanbul, İzmir ve Manisa illerinin değişik sağlık merkezlerinde soyutlanan 81 ÇİD-TB izolatında, yaygın ilaç direnç varlığının araştırılması ile Marmara ve Ege Bölgelerinde YİD-TB insidansının belirlenmesine yönelik çalışmalara katkı sağlanması amaçlanmış; aynı zamanda klinisyenlere ÇİD-TB hastalarının tedavisinde ikinci seçenek anti- TB ilaçların seçimi konusunda faydalanabilecekleri epidemiyolojik verilerin sunulması hedeflenmiştir. İzolatların birinci ve ikinci seçenek anti-TB ilaçlara duyarlılık durumları, tam otomatize florometrik BACTEC MGIT 960 cihazında (Becton Dickinson, ABD) modifiye Middlebrook 7H9 sıvı besiyeri kullanılarak araştırılmıştır. Çalışmaya dahil edilen 81 ÇİD-TB izolatının 43 (%53.1)’ü İstanbul, 26 (%32.1)’sı İzmir ve 12 (%14.8)’si Manisa ilinde yaşayan hastalardan soyutlanmıştır. Çalışma sonucunda YİD-TB tanımına uyan izolat saptanmamıştır. ÇİD-TB izolatlarının ikinci seçenek anti-TB ilaçlara direnç oranları, amikasin ve kanamisin için %1.2, ofloksasin ve levofloksasin için %2.5, kapreomisin için %14.8, etiyonamid için %37 olarak belirlenmiş, linezolide ise direnç tespit edilmemiştir. Levofloksasin-ofloksasin (p< 0.01), amikasin- kanamisin (p= 0.01) ve streptomisin-etiyonamid (p= 0.04) ilaçları arasında direnç durumları bakımından istatistiksel olarak anlamlı düzeyde uyum bulunduğu belirlenmiştir. Çalışmada, ÇİD-TB izolatlarında YİD-TB’nin görülmediği, etiyonamid ve kapreomisine karşı direncin yüksek bulunduğu, ÇİD-TB tedavisinde parenteral aminoglikozid grubundan amikasin veya kanamisinle birlikte, kinolonların ve yeterli klinik çalışmalar sonrası linezolidin güvenle kullanılabileceği sonucuna ulaşılmıştır.Increasing number of drug resistant tuberculosis (TB) cases, observed in recent years, is an important public health problem. Extensively drug resistant TB (XDR-TB) is the development of resistance against any fluoroquinolones and at least one of the injectable second line anti-TB drugs in addition to resistance against isoniazide and rifampicin which are the first line anti-TB drugs [definition of multidrug resistant TB (MDR-TB)]. Anti-TB therapy failed with first-line anti-TB drugs due to MDR-TB cases is being planned according to second-line anti-TB drug susceptibility test results if available and if not, standart treatment protocols are used. Although it is recommended that individual anti-TB therapy should be designed according to the isolate’s susceptibility test results, standart therapeutic protocols are always needed since second-line anti-TB drug susceptibility testing generally could not be performed in developing countries like Turkey. For this reason, nationwide and regional surveillance studies to determine the resistance patterns are always needed to make decisions about the standard therapy algorithms. In this study, it was aimed to investigate the presence of extensive drug resistance among 81 MDR-TB isolates obtained from various health care facilities from Istanbul, Izmir and Manisa and to determine the XDR-TB incidence in Marmara and Aegean regions. Furthermore, we aimed to provide epidemiological data to clinicians to support their choice of second-line anti-TB drugs for MDR-TB infections. Susceptibility testing of isolates for the first and the second-line anti-TB drugs were performed by using modified Middlebrook 7H9 broth in fluorometric BACTEC MGIT 960 system (Becton Dickinson, USA). Eighty-one MDR-TB isolates included in this study were isolated from 43 (53.1%) patients residing in Istanbul, 26 (32.1%) in Izmir and 12 (14.8%) in Manisa provinces. We could not find any isolate consistent with XDR-TB definition in this study. Second-line drug resistance rates of MDR-TB isolates to amikacin and kanamycin were 1.2%, ofloxacin and levofloxacin were 2.5%, capreomycin was 14.8%, ethionamide was 37% whereas linezolid resistance was not detected. Statistically significant correlation was detected between resistance rates of these antibiotic pairs; levofloxacin-ofloxacin (p&lt; 0.01), amikacin-kanamycin (p= 0.01) and streptomycin-ethionamide (p= 0.04). In our study, extensive drug resistance was not encountered in any MDR-TB isolates while high resistance rates was observed against ethionamide and capreomycin. It can be concluded that parenteral aminoglycosides amikasin and kanamycin, fluoroquinolones and linezolid seemed to be reliable anti-TB agents in MDR-TB treatment, however, further larger scale studies are needed

Glycopeptide Resistant Enterococcus faecium Isolated from Blood Culture

Glycopeptide resistant Enterococcus faecium strain was isolated from the blood cultures of a patient of the Haemotology Department of Gülhane Military Medical Academy, who was diagnosed acute myelocytic leukaemia. The strain was identified by the conventional and commercial automatic systems as E. faecium. Susceptibility pattern showed multiply resistance to all antibiotics except ciprofloxacin and levofloxacin. Using the E-test method, the MIC levels for vancomycin, teicoplanin, ciprofloxacin and levofloxacin were determined as 256 µg/mL, 64 µg/mL, 0.75 µg/mL and 1.5 µg/mL, respectively. VanA-1 and VanA-2 type resistance genes were detected by PCR. This strain is the first glycopeptide resistant E. faecium isolated from our hospital and Ankara and the second from Turkey. In the epidemiologic point of view, taking preventive measures against the wide spread of this dangereous bacteria is crucial

Plasmid-Mediated Quinolone Resistance Genes in Escherichia coli Urinary Isolates from Two Teaching Hospitals in Turkey: Coexistence of TEM, SHV, CTX-M and VEB-1 Type β-lactamases

Purpose: To evaluate the occurrence of plasmid-mediated quinolone resistance (PMQR) genes and the prevalence of extended spectrum β-lactamase (ESBL) types in Escherichia coli clinical isolates. Methods: Sixty-one ESBL-producing urinary E. coli isolates were studied. An antibiotic susceptibility test was performed using the disc diffusion method. ESBL production was determined using a double-disc synergy test for all isolates; E-test and Vitek 2 were used for plasmid-mediated quinolone resistance (PMQR)-positive isolates and their transconjugants. The presence of PMQR and β-lactamase genes was determined by polymerase chain reaction (PCR). Results: The strains displayed high rates of resistance to norfloxacin (80 %). The most frequent PMQR gene was aac(6’)-Ib-cr (45.9 %). In all, one qnrA1 (1.6 %), one qnrS1 (1.6 %), and two qepA1-positive isolates (5.7 %) were identified. The genes, qnrS1+aac(6’)-Ib-cr and qepA1, were co-expressed with blaCTX-M-15 gene, while qnrA1 occurred with blaTEM-1, blaSHV, and blaVEB-1 genes. The most frequent β-lactamase type was cefotaximase (CTX-M), which generally hydrolyzes cefotaxime (92 %) more than it does ceftazidime; followed by temoneira (TEM, 39 %); sulfhydryl variable (SHV, 5 %), and Vietnamese extended-spectrum beta– lactamase (VEB, 1.6 %). Conclusion: A high prevalence of aac(6’)-Ib-cr and CTX-M type β-lactamase was detected in ESBLproducing E. coli strains. This study also identified the co-expression of qnrA1 and blaVEB-1 genes and of qnrS1+aac(6’)-Ib-cr in E. coli isolates. The co-existence of PMQR genes with ESBLs may lead to a serious public health problem