Ahmet Pinar

Sandfly fever virus activity in central/northern Anatolia, Turkey: first report of Toscana virus infections.

Sandfly fever viruses (SFVs) cause febrile diseases as well as aseptic meningitis/encephalitis and include serotypes sandfly fever Sicilian virus (SFSV), sandfly fever Naples virus (SFNV) and Toscana virus (TOSV). Infections are endemic in the Mediterranean basin and data on SFV activity in Turkey are limited. In this study, sera from 1533 blood donors from the Ankara, Konya, Eskisehir and Zonguldak provinces of Turkey were evaluated for SFV exposure by indirect immunofluorescence test (IIFT) and confirmed by virus neutralization test (VNT). One hundred and two patients with central nervous system (CNS) infections of unknown aetiology were also tested via IIFT and real-time reverse-transcription PCR for SFV/TOSV. Rate of overall IgG reactivity in IIFT was 32.9% (505/1533) among blood donors. TOSV exposure was confirmed by VNT in all study regions. Exposure to the recently-identified serotype sandfly fever Turkish virus, as evaluated by VNT, was revealed in Konya and Ankara. SFNV exposure was identified in Konya and SFSV was observed to be present in all regions except Zonguldak. TOSV RNA was detected in 15.7% (16/102) and was accompanied by TOSV IgM in 25% (4/16) of the patients. Partial L and S sequences suggested that TOSV circulating in Turkey can be grouped into TOSV genotype A strains. Exposure to TOSV and other SFV serotypes was revealed in blood donors and CNS infections by TOSV were identified for the first time in Turkey. Infections are observed to be endemic in central Anatolia and should be considered as aetiologic agents in cases/outbreaks of fever and meningoencephalitis.

Investigation of HSV-1, HSV-2, CMV, HHV-6 and HHV-8 DNA by real-time PCR in surgical resection materials of epilepsy patients with mesial temporal lobe sclerosis

OBJECTIVE The objective of this study is to investigate the presence of viral DNAs of HSV-1, HSV-2, HHV-6, HHV-8, and CMV in hippocampus of the patients with mesial temporal lobe epilepsy (MTLE) syndrome. METHODS Pathological specimens were obtained from 33 patients with MTLE undergone temporal lobectomy with amygdalo-hippocampectomy due to intractable seizures. Autopsy materials from the hippocampus of 7 patients without neurological disease were used as controls. The data was also correlated with the clinical history of patients including febrile convulsions, age, and history of CNS infections. Real-time polymerase chain reaction method was performed for detection of DNAs of these viruses. RESULTS HHV-6, HSV-1 and HHV-8 were detected in the hippocampus of 3, 2 and 1 patients with MTLE respectively. None of the hippocampus of patients with MTLE was positive for DNA of HSV-2 and/or CMV. Three patients with positive HHV-6 DNAs had febrile convulsions and family history for epilepsy. None of our control specimens showed PCR positivity to any of the 5 tested viruses. CONCLUSIONS Our study is the first to report the presence of HHV-8 viral genome in the brain tissue of patient with MTLE. Viral DNAs were detected in a total of 18% of the patients in this study; we can conclude that activity of the latent virus in patients with hippocampal sclerosis should be more extensively studied to establish its role in active infection.

Identifying the etiologic role of Parvovirus B19 in non-immune hydrops fetalis by histopathology, immunohistochemistry and nucleic acid testing: a retrospective study

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).

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.

Serotype Prediction for Frequently Isolated Serotypes of Streptococcus Pneumoniae by Heteroduplex Analysis and Modification of This Technique to Real-Time Fluorometric Nucleic Acid Detection System

ABS TRACT Ob jec ti ve: Se ve ral mo le cu lar se roty pe pre dic ti on met hods ha ve be en pub lis hed in or der to simp lify the pne u mo coc cal se roty ping. The aim of this study is to de ve lop a mo le cu lar se roty pe pre dic ti on tech ni qu e al ter na ti ve to ot her mo le cu lar ap pro ac hes. Ma te ri al and Met hods: S. pne u mo ni a e se roty pes 1, 3, 5, 6A, 7F, 8, 9V, 11A, 14, 15B, 18C, 19A, 19F and 23B, that are the most fre qu ently iso la ted se roty pes from in fec ti ons in Tur key, we re inc lu ded in the study. S. pne u mo ni a e cpsA and cpsB ge nes that are in vol ved in the pro ces sing, re gu la ti on and ex port of the cap su lar poly sacc ha ri des, we re amp li fi ed by polymerase chain reacsion (PCR). Then, he te ro dup lex analy sis was per for med to the 1.8 kb PCR pro ducts ob ta i ned from all se roty pes with cons tant se roty pe 1. This tech ni qu e was then mo di fi ed to re al-ti me flu o ro met ric nuc le ic acid de tec ti on system. Re sults: The PCR pro ducts ob ta i ned from se roty pes 1, 6A, 14 and 19F sho wed only ho mo dup lex bands whi le tho se from se roty pes 3, 5, 7F, 8, 9V, 11A, 15B and 23B sho wed spe ci fic he te ro dup lex bands. Alt ho ugh se roty pes 18C and 19A sho wed a spe ci fic he te ro dup lex ban ding pat tern, they co uld not be dif fe ren ti a ted from each ot her. When furt her he te ro dup lex analy sis was per for med to se roty pes 1, 6A, 14 and 19F with se roty pe 5 as a cons tant com po nent, se roty pe 1 and 6A co uld be dif fe ren ti a ted from the ot hers. He te ro dup lex and ho mo dup lex DNA strands co uld be dis tin gu is hed by mel ting cur ve analy sis in re al-ti me flu o ro met ric nuc le ic acid de tec ti on system. Conc lu si on: As a ra pid and cost-ef fec ti ve met hod, re al-ti me he te ro dup lex analy sis may be an al ter na ti ve to ot her mo le cu lar met hods for se roty pe pre dic ti on of pne u mo coc ci.