Molecular characterization of Mycobacterium tuberculosis isolates from Izmir, Turkey.

In recent years, molecular typing methods have been used in epidemiologic studies of Mycobacterium tuberculosis isolates in various areas of the world. However, there have been few data on this issue in Turkey. We describe the molecular characterization of 56 Mycobacterium tuberculosis isolates recovered from individual patients in Izmir and the surrounding area by three different molecular methods. Isolated M. tuberculosis strains were characterized by IS6110 RFLP, spoligotyping and major genetic group designation. In total, 51 RFLP and 35 spoligopatterns were identified. Fourteen (25%) isolates were indicated as low copy number. Based on three genotypic characterization methods together, five clusters with two isolates each were identified. Most of the isolates (98.2%) were assigned as genetic groups 2 or 3. Only one isolate was identified as Beijing family strain (principal genetic group 1). The shared international clades were found to be Beijing-family, var T1 (ST 37), LAM (Latin-American-Mediterranean) 7 (ST 41), LAM 9 (ST 42), Haarlem 1 (ST 47), Haarlem 3 (ST 50) and T1 (ST 53). In this study, IS6110 RFLP, spoligotyping and major genetic group designation were found to be useful methods for molecular epidemiologic studies.

Effect of subminimal inhibitory concentrations of three fluoroquinolones on adherence of uropathogenic strains of Escherichia coli.

The effect of subinhibitory concentrations (1/2-1/32 x MIC) of ciprofloxacin, ofloxacin and levofloxacin on the adherence of three strains of Escherichia coli (a mannose-resistant haemagglutinating clinical isolate, a non-haemagglutinating clinical isolate and the mannose-resistant haemagglutinating ATCC 25922 strain) were studied. Ciprofloxacin had the lowest MIC values but only the 1/2 MIC concentration inhibited adherence of mannose-resistant haemagglutinating strains after exposure to subMIC values. Significant inhibition of adherence was observed with 1/4 x MIC ofloxacin for both haemagglutinating isolate (27096) and the ATCC strain. Levofloxacin might be more effective and safer than ciprofloxacin and ofloxacin as a long acting fluoroquinolone at subMIC values in patients with UTI.

[Comparison of routine sensitivity tests and mecA gene analysis for determination of methicillin resistance in Staphylococcus aureus isolates]. / Staphylococcus aureus izolatlarinin metisilin direncinin saptanmasinda rutin duyarlilik testleri ve mecA gen analizi sonuçlarinin karsilastirilmasi.

Currently, oxacillin disk diffusion test is the most frequently employed method to detect the methicillin resistance of Staphylococcus aureus isolates. However, due to some of the test conditions, errors may occur during the detection of heteroresistant bacteria. It is now widely accepted that lower incubation temperatures (< or = 35 degrees C) and media with a NaCl concentration of 2-4%, could facilitate detection. In our study, methicillin (oxacillin) susceptibilities of 125 S. aureus isolates were determined by the disk diffusion and microdilution tests as defined by National Committee for Clinical Laboratory Standards (NCCLS), and the results were compared with those of mecA gene analysis. In the routine susceptibility tests, 75 isolates were found to be methicillin resistant (MRSA), whereas 50 were found to be susceptible (MSSA). Various induction tests were performed to investigate the heterogeneous resistance among methicillin-susceptible isolates. These induction tests showed that the MIC values of seven isolates reached to the resistant levels, therefore these isolates should be accepted as "borderline oxacillin resistant S. aureus" isolates lacking the mecA gene. The susceptibility tests and mecA gene analysis of the remaining isolates yielded compatible results. In conclusion, susceptibility tests, when performed according to NCCLS recommendations, are found to be reliable and decisive, for the detection of methicillin resistance of S. aureus.

[Comparison of indirect immunofluorescence and enzyme immunoassay methods for the determination of antinuclear antibodies]. / Antinükleer antikorlarin saptanmasinda indirek immünofloresan ve enzim immün yöntemlerinin karsilastirilmasi.

Antinuclear antibodies (ANA) are widely used for screening and monitoring of connective tissue diseases (CTD). Indirect immunofluorescence assay (IFA) is the standard method which is more often preferred for detecting these antibodies. Another method is enzyme immunoassay (ELISA) which includes extractable nuclear antigens (ENA). The aim of this study was to compare two different methods in view of their performances in the detection of ANA. A total of 27 sera from patients prediagnosed as different types of CTD, were screened by ANA-IFA (Zeus Scientific Inc, USA) and ELISA (Zeus Scientific Inc, ENA Profile-6, USA) methods. In addition, specific staining patterns of ANA on HEp-2 cells as a substrate, were enrolled with IFA. As a result, ANA positivity was detected in all of the 27 samples (100%) by IFA, and only in 7 (25.9%) by ELISA. The concordance rate between two different assays was estimated as 38.6%, and a statistically significant difference was found between the methods in the detection of ANA (x2=20, p<0.001). In conclusion, ANA-IFA method is still a reliable routine screening method for the laboratory diagnosis of CTD, while ENA Profile-6 ELISA may give false negative results, because of its limited antigen content, and should be supported with additional antigens.

Species distribution and antifungal susceptibilities of dermatophytes during a one year period at a university hospital in Turkey.

Dermatophyte infections have been considered to be a major public health problem in many parts of the world. The aim of this study was to determine the causative agents of dermatophytoses and their antifungal susceptibilities in a Turkish University Hospital, west of Turkey. A total of 926 patients suspected to have dermatophytic lesions were examined over a period of 1 year (2001-2002). Samples collected from skin, hair and nails were submitted to direct microscopical examination using KOH and Calcofluor white stain, cultured on Sabouraud dextrose agar and Mycosel agar. The prevalence of dermatophytoses was 7.34% (68/926). Trichophyton rubrum was the most frequent dermatophyte isolated (56%) followed by T. mentagrophytes (38%), T. violaceum (1.5%), T. verrucosum (1.5%), Microsporum canis (1.5%) and Epidermophyton floccosum (1.5%). Tinea pedis (47%) was the most common type of infection, followed by tinea unguium (29%), tinea inguinalis (15%), tinea corporis (7.4%) and tinea capitis (1.6%). Secondary, we have tested 68 strains of dermatophytes against four antifungal agents following mainly the National Committee for Clinical Laboratory Standards M38-P standard for filamentous fungi. In general, all antifungals were shown to be highly effective and itraconazole and naftifine appeared more active than ketoconazole and oxiconazole.

[The importance of antinuclear (ABA) and anti-double stranded dna (anti-dsDNA) antibodies in the diagnosis of connective tissue diseases.]. / Konnektif doku hastaliklarinin tanisinda antinükleer (ANA) ve anti-double stranded dna (anti-dsDNA) antikorlarinin önemi.

Connective tissue diseases (CTD) are characterized by the presence of autoantibodies against several tissues. These autoantibodies occur against cell membrane, cell receptors, plasma proteins, and cytoplasmic and nuclear components. In laboratories, anti-nuclear antibodies (ANA) and anti-double stranded DNA (anti-dsDNA) antibodies are widely used in diagnosis of CTD. The aim of this study was to investigate the presence and accompaniment of ANA and anti-dsDNA antibodies in diagnosis of several CTD and also to study the prevalence of ANA and anti-dsDNA in a group of 88 patients with various types of CTD. ANA were detected by immunofluorescence (IFA) using HEp-2 cells (Zeus Scientific, Inc. USA) and anti-dsDNA antibodies using Crithidia luciliae (BioSystems, Spain) as substrates in immunofluorescence. ANA Western Blot (WB) Immunoassay (ImmuBlot, International Immuno-Diagnostics, USA) was also used along with the tests referred to previously. ANA was found in the sera of 84 (96.5%) patients while anti-dsDNA was detected in 7 (7.95%). Moreover different fluorescence patterns were also evaluated with ANA IFA in accordance with anti-dsDNA results. Mixed patterns in three and a homogeneous pattern in four anti-dsDNA positive patients' sera were determined on HEp-2 cell line by IFA. Seven sera which were ANA and anti-dsDNA positive with IFA were also found to be positive with WB and their ANA patterns with the specific ANA WB bands were also evaluated. It was observed that IFA results were in concordance with WB results. Our data indicated that the above findings should be controlled and evaluated with a more advanced method such as western blotting technique in order to confirm the presence of specific antibodies along with clinical outcome of the patients. As a result we think that ANA WB method is an appropriate technique in diagnosis of CTD as anti-dsDNA and ANA bands can be evaluated together with this method.