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1
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Performance of three PCR methods targeting different regions of viral genome for the detection of TTV in Non A-E hepatitis, chronic B and C hepatitis and healthy blood donors

100%
Ergunay K., Sivri B., Karabulut E., Ustacelebi S., Bayraktar Y.
Open Medicine
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2006
|
vol. 1
|
issue 3
250-260
EN
TT virus (TTV) was suggested to be the etiologic agent for non A-E hepatitis but this could not yet be proven due to high detection rates not only in hepatitis but also in healthy persons and sensitivity differences of PCR methods employed. The aim of this study was to evaluate TTV DNA positivity in non A-E hepatitis cases, chronic HBV and HCV hepatitis cases and healthy blood donors via PCR systems that target all regions of the viral genome used for viral detection. 23 non A-E hepatitis, 28 chronic HCV, 21 chronic HBV cases and 56 healthy blood donors were included in the study and evaluated by PCR protocols that target 5′-UTR, 3′-UTR and N22 (ORF1) regions. As a result, 3′-UTR and 5′-UTR PCR had comparable detection rates that were higher than N22 PCR. Differences in detection rates among study groups were not statistically significant for any PCR method. Hepatic enzyme levels of the patients were not correlated with the presence of TTV DNA. Detection rate was significantly higher for Non A-E hepatitis group when positivity rates from all methods were combined. These results suggest an alteration of viral genotypes in Non A-E hepatitis which might be associated with pathogenesis.
2
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Identifying the etiologic role of Parvovirus B19 in non-immune hydrops fetalis by histopathology, immunohistochemistry and nucleic acid testing: a retrospective study

86%
Ergunay K., Altinok G., Gurel B., Pinar A., Sungur A., Balci S., Ustacelebi S.
Open Medicine
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2007
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vol. 2
|
issue 3
271-279
EN
Intrauterine Parvovirus B19 infections may cause fetal anemia, non-immune hydrops fetalis or abortion. This study focuses on the pathogenic role of Parvovirus B19 in non-immune hydrops fetalis at Hacettepe University, a major reference hospital in Turkey. Twenty-two cases of non-immune hydrops fetalis were retrospectively selected out of a total of 431 hydrops fetalis specimens from the Department of Pathology archieves. Paraffine embedded tissue sections from placental and liver tissues from each case were evaluated by histopathology, immunohistochemistry, nested PCR and commercial quantitative Real-time PCR. Viral DNA was detected in placental tissues by Real-time PCR in 2 cases (2/22, 9.1%) where histopathology also revealed changes suggestive of Parvovirus B19 infection. No significant histopathologic changes were observed for the remaining sections. Nested PCR that targets the VP1 region of the viral genome and immunohistochemistry for viral capsid antigens were negative for all cases. As a result, Parvovirus B19 is identified as the etiologic agent for the development of non-immune hydrops fetalis for 9.1% of the cases in Hacettepe University, Turkey. Real-time PCR is observed to be an effective diagnostic tool for nucleic acid detection from paraffine embedded tissues. Part of this study was presented as a poster at XIIIth International Congress of Virology, San Francisco, USA (Abstract V-572).
3
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Comparason of extraction methods for PCR detection of Burkholderia cepacia complex (BCC) from cystic fibrosis patients

86%
Ergunay K., Yurdakul P., Sener B., Ozcelik U., Karabulut E., Kiper N.
Open Medicine
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2008
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vol. 3
|
issue 2
157-162
EN
Direct detection of Burkholderia cepacia complex (BCC) and its genomovars from sputum by molecular tests emerges as a method for rapid identification. In this study, four DNA extraction methods were evaluated for the identification for BCC from sputum of CF patients. Sputa from 28 CF patients were aliquoted and spiked with BCC reference strain. Boiling, phenol-chloroform, CTAB methods and a commercial spin column kit was used for DNA extraction. Total DNA yields were determined by spectrophotometry and single-round recA PCR was used for detection of BCC. No significant difference was observed in DNA yields from different extraction methods. Lower limit of detection for recA PCR was determined as 106 cfu/ml. Amplification was observed in 7/16 (43.7%) of sputa for boiling, 8/16 (50%) of sputa for CTAB and 13/16 (81.2%) of sputa for phenol-chloroform method and spin column kit in the assay sensitivity range determined in the study. Phenol-chloroform and commercial spin column kit were found to be better suited for DNA purification from sputum of CF patients for BCC identification. Diagnostic impact of single-round recA PCR directly from sputum was limited to chronically-infected patients.
4
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Helicobacter pylori isolation, serology and cagA, cagE and virB11 detection in patients with non-ulcer dyspepsia from Turkey: Correlation with histopathologic findings

86%
Ilga U., Ozyurt M., Yildirim S., Ergunay K., Ardic N., Demirturk L., Haznedaroglu T.
Open Medicine
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2008
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vol. 3
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issue 1
41-46
EN
Colonization with Helicobacter pylori (HP) may have major clinical consequences and HP virulence factors are associated with more severe gastroduodenal pathologies. In this study, prevalence of HP in patients with Non-Ulcer Dyspepsia (NUD) was determined by rapid urease test and culture and correlations of histopathologic changes with bacterial virulence factors and serologic profiles were investigated. Gastric biopsies from sixty-nine patients admitted to Haydarpasa Training Hospital Department of Gastroenterology were evaluated for rapid urease, HP isolation and examined histopathologically. PCR was employed for HP confirmation and detection of HP cagA, cagE and virB11 genes. For each patient, IgG and IgA antibodies and anti-cagA antibodies were also determined by ELISA tests. HP was isolated and confirmed by PCR in 74% (51/69) of the patients. Anti-HP IgG and IgA were detected in 96% (49/51) and 53% (27/51), respectively. Anti-cagA were present in 51% (26/51). cagA, cagE and virB11 were positive in 56.8% (29/51), 60.7% (31/51) and 58.8% (30/51) of the patients, respectively. Statistically significant correlation was observed between cagA PCR and inflammation/activity scores. Detection of cagA by molecular assays can be an alternative test that can be employed for individual patient assessment.
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