Detection and molecular characterisation of adenovirus in children under 5 years old with diarrhoea

Background/aim: To determine the frequency, genotype distribution, and genetic relatedness of adenoviruses in children under 5 years old with diarrhoea and to investigate their distribution according to clinical findings, age, months, and seasons. Materials and methods: Stool samples were collected from 180 children with acute gastroenteritis who presented from July 2007 through June 2011 at the Ankara Training and Education Hospital. Stool samples were analysed by immune chromatographic test (ICT), enzyme immunoassay (EIA), and polymerase chain reaction (PCR). All adenovirus types were determined by nucleotide sequence analysis. A phylogenetic tree was constructed by Mega 6.0 using the neighbour-joining method. Results: Five percent of the samples were positive for adenovirus (9/180) by ICT, 6.1% (11/180) by EIA, and 13.9% (25/180) by PCR. Adenovirus gastroenteritis did not show any differences in age group, sex, month, or season. In this study, 16 (64%) of the PCR positive samples were AdV41, 6 (24%) were AdV40, 2 (8%) were AdV31, and 1 (4%) was AdV7, as determined by nucleotide sequencing. Conclusion: AdV31 and AdV7 were associated with gastroenteritis. Adenovirus serotypes showed a similarity of 80% (20/25) and 20% (5/25) with Asian and American serotypes, respectively.

HPV types and E6/E7 mRNA expression in cervical samples from Turkish women with abnormal cytology in Ankara, Turkey.

BACKGROUND/AIM: Human papillomaviruses have been established as a risk factor for invasive carcinoma of the uterine cervix. HPV E6/E7 oncogene expression has recently emerged as a promising biomarker to determine the risk for progression to high-grade cervical lesions. The aim of this study was to evaluate HPV mRNA and DNA detection in samples with abnormal cytology. MATERIALS AND METHODS: Cervical specimens were obtained at the Department of Obstetrics and Gynecology via cervical brushes during January-October 2011. Liquid-based cytology slides were evaluated according to the 2001 Bethesda System. Cytology specimens from a total of 81 women with abnormal cytology were included. Real-time PCR and NASBA assays were performed to detect HPV DNA and E6/E7 mRNA, respectively. RESULTS: HPV DNA was identified in 73 samples (90.1%). HPV E6/E7 mRNA expression was observed in 45 samples (55.6%). A statistically significant difference was observed among cytological diagnosis groups. In 25 patients, a biopsy was performed during the follow-up. HPV DNA was detected in all of these patients. HPV E6/E7 expression was present only in CIN I-III diagnosed patients. CONCLUSION: The E6/E7 mRNA test is a robust indicator of cytological atypia and correlates better with progressive lesions than DNA assays.

Re-emergence of genotype G9 during a five-and-a-half-year period in Turkish children with rotavirus diarrhea.

This study was done to understand the dynamics of rotavirus genotype distribution in Turkish children. Samples were collected from January 2006 through August 2011 from children at a hospital in Ankara. Rotavirus was detected in 28 % (241/889) of the samples. Genotype G9P[8] was predominant (28 %), followed by G1P[8] (16.3 %) and G2P[8] (15.9 %). G9 was absent in the samples from 2006 and 2007 and then re-emerged in 2008 and increased gradually. Phylogenetic analysis showed that Turkish G9 rotaviruses of the present study formed a sublineage with strains from Italy and Ethiopia, possibly indicating spread of a clone in these countries.

Microbiologic Examination of Bandage Contact Lenses Used after Corneal Collagen Cross-linking Treatment.

PURPOSE: To investigate the agents of bacterial contamination of contact lenses after corneal collagen cross-linking (CCL), and to present the possible changes of ocular flora after riboflavin/ultraviolet A. METHODS: Seventy-two contact lenses of patients who underwent CCL and 41 contact lenses of patients who underwent photorefractive keratectomy (PRK) as control group were enrolled to the study. After 48 h of incubation, broth culture media was transferred to plates. Samples were accepted as positive if one or more colony-forming units were shown. RESULTS: There were positive cultures in 12 (16.7%) contact lenses in the CCL group and 5 (12.2%) had positive cultures in PRK group. Coagulase-negative staphlycocci (CNS) were the most frequent microorganism. Alpha hemolytic streptococci and Diphteroid spp. were the other isolated microorganisms. CONCLUSIONS: Bacterial colonization can occur during and early after the CCL procedure in epithelial healing. To prevent corneal infections after the treatment, prophylactic antibiotics should be prescribed.

[Using Galleria mellonella as an in vivo model to study the virulence of some bacterial and fungal agents]. / Bazi bakteri ve mantarlarin virülansinin arastirilmasinda Galleria mellonella'nin in vivo model olarak kullanilmasi.

Non-vertebrate hosts, such as Galleria mellonella, namely wax moth, have been used to study microbial virulence and host defense. This organism has advantages as it is economical, ethically expedient and easy to handle. Here we describe an experimental in vivo study using the larvae of Galleria mellonella infected with some bacterial and fungal pathogens. In this study, extended-spectrum beta-lactamase (ESBL) producing and non-producing Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa, colistin resistant and susceptible Acinetobacter baumanii clinical strains; Candida albicans (ATCC 10231), Scedosporium aurantiacum (CBS 136047) and Pseudallescheria boydii (CBS 117410) reference strains, and Aspergillus terreus and Fusarium oxysporum clinical strains were used as pathogens. The larvae of G.mellonella were challenged with these bacterial and fungal strains, and the mortality rates were calculated using Kaplan-Meier plots. Mortality rates at 16th hour were found as 83% for the larvae infected with both ESBL positive and negative E.coli, ESBL negative K.pneumoniae and ESBL positive P.aeruginosa; 91% for ESBL positive K.pneumoniae; 75% for ESBL negative P.aeruginosa; 66% for both colistin resistant and susceptible A.baumanii strains. All larvae infected with bacteria died within the first 24 hour. Larvae infected with bacteria showed significantly higher mortality rates than those infected with fungi. Mortality rates at 16th hour were found as 0% for C.albicans and F.oxysporum, 16% for S.aurantiacum, 8% for P.boydii and A.terreus; at 24th hour that was 25% for C.albicans and P.boydii, 33% for S.aurantiacum, A.terreus and F.oxysporum; at 48th hour that was 33% for C.albicans, 50% for P.boydii and F.oxysporum, 58% for A.terreus, and 66% for S.aurantiacum; in 72 hours that was 58% for C.albicans and F.oxysporum, 66% for P.boydii, 75% for A.terreus and S.aurantiacum, in 96 hours that was 83% for C.albicans, P.boydii and F.oxysporum, 91% for A.terreus and S.aurantiacum. As a result of this study, potential evidences provided that bacteria were more virulent than fungi for G.mellonella larvae model, each fungal species showed different virulence patterns, and bacterial virulence was correlated neither with species nor antibiotic susceptibility.

[Investigation of norovirus infection incidence among 0-5 years old children with acute gastroenteritis admitted to two different hospitals in ankara, Turkey]. / Akut Gastroenterit Nedeniyle Ankara'da Iki Farkli Hastaneye Basvuran 0-5 Yas Arasi Çocuklarda Norovirus Enfeksiyonu Sikliginin Arastirilmasi

Norovirus causes severe gastroenteritis requiring hospitalization especially in children less than five years of age both in developed and developing countries. Therefore, we aimed to investigate the incidence of norovirus (NoV) in 0-5 years old children with acute gastroenteritis in two large hospitals in Ankara, Turkey. Stool samples were obtained from 1000 (413 female, 587 male) children between 0-5 years old with acute gastroenteritis who attended to the Department of Paediatrics, Ministry of Health Ankara Training and Education Hospital and affiliated hospital of Gazi University Faculty of Medicine between October 2004 and June 2011. Antigens of norovirus GI and GII genogroups in the stool specimens were detected by ELISA (RIDASCREEN® Norovirus (C1401) 3rd Generation, R-Biopharm, Germany). Norovirus GI and GII antigens were determined in a total of 141 (14.1%) samples, of them 62 (15%) were female and 79 (13.5%) were male, yielding no statistically significant difference (p> 0.05). The highest NoV positivity was detected in children between 12-23 months (17.1%), however there was no statistically significant difference between ELISA positivity and age (p> 0.05). NoV detection rate was highest in 2007 (18.4%) and in 2009 (18%), and the difference regarding ELISA positivity among the study years was not statistically significant (p> 0.05). The prevalences of norovirus infection in spring, summer, autumn and winter were 13.8%, 17.7%, 14.7% and 11.2%, respectively. Therefore no seasonal variation was found in the incidence of norovirus infection. However when the monthly prevalence was analyzed, a statistically significant difference was found (p< 0.05) between the rate of norovirus infection in july (24.2%) and december (4.1%). When evaluating the clinical symptoms, all of 141 patients (100%) had diarrhoea, while 72 (51.1%) had vomiting. Stool samples were also evaluated for the presence of parasitic and bacterial agents. Coinfection rate with parasites was detected as 3.3% (4/122; norovirus + Entamoeba histolytica in three cases, norovirus + Enterobius vermicularis in one case), while no pathogenic bacteria were isolated from norovirus positive stool samples. The prevalence rate of 14.1% for NoV GI/GII infection detected in this retrospective study including 0-5 years old children in Ankara for 2004-2011 period was thought to reflect the regional data and would contribute to national epidemiological data. We anticipate that the incidence of norovirus will increase in 0-5 year old children as a result of increasing use of rotavirus vaccine in Turkish children. It was concluded that, NoV antigen detection tests should be used in routine laboratories for appropriate diagnosis of sporadic and/or epidemic norovirus infections.