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1

Isolation of Acanthamoeba isolates belonging to T2, T3, T4 and T7 genotypes from environmental samples in Ankara, Turkey

100%
Kilic A., Tanyuksel M., Sissons J., Jayasekera S., Khan N. A.
Acta Parasitologica
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2004
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tom 49
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nr 3
Acanthamoeba keratitis is a blinding infection that is becoming increasingly important in human health. Early diagnosis is a prerequisite for successful treatment and requires identification of Acanthamoeba at the genotypic level. The genus Acanthamoeba consists of both pathogenic and non-pathogenic species and has been recently classified into 13 different genotypes, T1-T12 and T14. More importantly, 95% of Acanthamoeba isolates that produce keratitis belong to T4 genotypes. In this study, we attempted to determine whether predominance of T4 isolates in Acanthamoeba keratitis is due to greater virulence or greater prevalence. We isolated 18 Acanthamoeba isolates from environmental samples in Ankara, Turkey and determined their pathogenic potential by means osmotolerance, temperature tolerance and in vitro cytotoxicity assays using corneal epithelial cells. Ribosomal DNA sequencing revealed that 10 isolates belong to T2, 5 belong to T3,2 belong to T4 and one belongs to T7 genotype. As expected, T3 and T4 isolates exhibited the most pathogenic traits and were osmotolerant, temperature tolerant and exhibited severe corneal epithelial cell cytotoxicity indicating their pathogenic potential. Overall these data indicate that high frequency of T4 isolates in keratitis cases may well be due to their greater virulence. This is the first report presenting environmental distribution of Acanthamoeba in Ankara, Turkey.
2

Simultaneous detection and differentiation of pathogenic and nonpathogenic Leptospira spp. by multiplex real-time PCR (TaqMan) assay

86%
Bedir O., Kilic A., Atabek E., Kuskucu A. M., Turhan V., Basustaoglu A. C.
Polish Journal of Microbiology
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2010
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tom 59
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nr 3
Leptospirosis, caused by pathogenic Leptospira, is one of the most important zoonoses in the world. Several molecular techniques have been developed for detection and differentiation between pathogenic and saprophytic Leptospira spp. The aim of this study was to develop a rapid and simple assay for specific detection and differentiation of pathogenic Leptospira spp. by multiplex real-time PCR (TaqMan) assay using primers and probes targeting Leptospira genus specific 16S ribosomal RNA gene, the pathogen specific ligA/B genes and non-pathogen Leptospira biflexa specific 23S ribosomal RNA gene. Sixteen reference strains of Leptospira spp. including pathogenic and nonpathogenic and ten other negative control bacterial strains were used in the study. While the 16S primers amplified target from both pathogenic and non-pathogenic leptospires, the ligA/B and the 23S primers amplified target DNA from pathogenic and non-pathogenic leptospires, respectively. The multiplex real-time PCR (TaqMan) assay detection limit, that is, the sensitivity was found approximately 1 x 10² cells/ml for ligA/B gene and 23S ribosomal RNA gene, and 10 cells/ml 16S ribosomal RNA. The reaction efficiencies were 83-105% with decision coefficients of more than 0.99 in all multiplex assays. The multiplex real-time PCR (TaqMan) assay yielded negative results with the ten other control bacteria. In conclusion, the developed multiplex real-time PCR (TaqMan) assay is highly useful for early diagnosis and differentiation between pathogenic and non-pathogenic leptospires in a reaction tube as having high sensitivity and specificity.
3

Macrolide-lincosamide-streptogramin B resistant phenotypes and genotypes for methicillin-resistant Staphylococcus aureus in Turkey, from 2003 to 2006

86%
Gul H. C., Kilic A., Guclu A. U., Bedir O., Orhon M., Basustaoglu C.
Polish Journal of Microbiology
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2008
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tom 57
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nr 4
Methicillin-resistant Staphylococcus aureus (MRSA) strains with inducible macrolide-lincosamide-streptogramin B (iMLSB) resistance phenotype may lead to clinical failure during clindamycin (CLI) therapy. The aim of this study was to determine the incidence of MLS|t phenotypes by using D-test method and genotypes by using multiplex real-time PCR method in MRSA strains. A total of 265 MRSA strains were obtained from clinical samples from hospitalized and outpatients. Of the MRSA isolates, 225 (84.9%) were resistant to erythromycin (ERT), and 170 (64.1%) to CLI. Among the 225 ERT-resistant MRSA strains, the constitutive MLS|f (cMLSu) rate was found in 49.3%, iMLSB in 39.1% and the M phenotype in 11.5%. Overall, ermA, ermC, ermA+ermC, msrA, ermC+msrA, and ermA+ermC+msrA genes were detected in 85 (37.7%), 60 (26.6%), 42 (18.6%), 26 (11.5%), 11 (4.8%), and 1 (0.4%) isolates, respectively. Most prevalent resistance determinant in MRSA strains was ermA", which was detected in 37.7% of the isolates. The 26 MRSA strains with M phenotype harboured only msrA gene. In conclusion, due to aware of the potential of CLI treatment failure, D-test should be performed and reported in MRSA strains in clinical laboratories. The multiplex real-time PCR method is easy to perform, fast and reliable method for the detection of MLSB resistance genotypes.
4

Detection of cytomegalovirus and Helicobacter pylori DNA in arterial walls with grade III atherosclerosis by PCR

86%
Kilic A., Onguru O., Tugcu H., Kilic S., Guney C., Bilge Y.
Polish Journal of Microbiology
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2006
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tom 55
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nr 4
It has been suggested that some microorganisms may play a role in the etiology or progression of atherosclerotic plaques. The purpose of this study was to assess for the presence of Helicobacter pylori and cytomegalovirus (CMV) DNA using polymerase chain reaction (PCR) technique in vascular-wall specimens obtained during autopsy. Four to 5 mm long samples from 3 different vascular wall specimens (coronary, carotid and abdominal aortas) of 30 patients (23 male, 7 female) were taken for pathologic and microbiologic investigations during autopsy. H. pylori DNA was found in 48.2% atherosclerotic and 19.6% non-atherosclerotic vascular wall specimens, whereas CMV DNA was found in 37.9% atherosclerotic and 32.7% non-atherosclerotic vascular wall specimens. In terms of CMV DNA detection, no statistically significant differences between the atherosclerotic and non-atherosclerotic groups were present (P>0.05). However, there was a statistically significant difference between the atherosclerosis and non-atherosclerotic groups in terms of H. pylori DNA in coronary and abdominal aorta arteries (p = 0.016 and p = 0.0029 respectively) but not in carotid arteries (p= 1.00). In conclusion, the correlation between H. pylori and atherosclerosis could be suggested. These finding warrant further investigation regarding the role of H. pylori in atherosclerosis.
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