[Show abstract][Hide abstract] ABSTRACT: Tick-borne encephalitis virus (TBEV) is the aetiological agent of tick-borne encephalitis (TBE), a potentially fatal central nervous system infection of humans. TBE is endemic in many areas of Europe and Asia; however, very scarce data on TBEV activity are available from Turkey. We aimed to identify TBEV exposure in healthy blood donors and the impact of TBEV in central nervous system infections in Central/Northern Anatolia. Two-thousand four hundred and fifty four sera, collected from blood donors at Ankara, Konya, Eskişehir and Zonguldak branches of the Turkish Red Crescent Middle Anatolia Regional Blood Center, were analysed for TBEV serosurveillance. Paired serum and cerebrospinal fluid samples from 108 patients with the diagnosis of aseptic meningitis/encephalitis of unknown aetiology were also evaluated to identify TBE and neuroborreliosis cases. Commercial enzyme-linked immunosorbent assays and indirect immunofluorescence tests were employed for antibody detection. Forty-seven donor samples (1.9%) were reactive for TBEV IgG. In 25 persons with IgG reactivity (53.1%), risk factors for tick-borne infections were revealed. One sample from Zonguldak province (1/198; 0.5%) in the Black Sea region of Turkey was confirmed to possess neutralizing antibodies via plaque reduction neutralization test. TBEV IgM was detected in 9.2% (8/108) of the patients. IgM was accompanied by IgG reactivity in two persons where, in one, recent history of a tick bite was also identified. Intrathecal antibody production for TBEV could not be demonstrated. No evidence for Borrelia infections could be found. Confirmed exposure to TBEV and/or an antigenically similar tick-borne flavivirus is documented for the first time in blood donors in Zonguldak in Northern Anatolia. Probable cases of TBE have also been identified from Central Anatolia. The epidemiology of TBEV activity in Turkey needs to be assessed and benefits of vaccination for general population, risk groups or travellers must be considered.
Zoonoses and Public Health 05/2011; 58(3):220-7. DOI:10.1111/j.1863-2378.2010.01342.x · 2.07 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Clin Microbiol Infect 2011; 17: 575–581
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.
[Show abstract][Hide abstract] ABSTRACT: Sandfly fever virus (SFV) serotypes sandfly fever Naples virus, sandfly fever Sicilian virus, and sandfly fever Cyprus virus cause febrile diseases, whereas Toscana virus (TOSV) is responsible for aseptic meningoencephalitis. Diagnosis and surveillance of TOSV depend heavily on virus serology, and various commercial assays utilizing various antigen sources and formats have been available. The aim of this study was to perform comparative evaluation of commercially available serological assays for anti-TOSV immunoglobulins.
A collection of 120 sera from healthy blood donors from an endemic region, previously identified to be reactive for antibodies against various SFV serotypes by indirect immunofluorescence test (IIFT), was reevaluated for IgG/IgM via IIFT, enzyme-linked immunosorbent assay, and an immunoblot assay manufactured by Euroimmun, Diesse, and Mikrogen, respectively. Virus neutralization test (VNT) was performed for 99 sera using standard TOSV, sandfly fever Sicilian virus, and sandfly fever Naples virus strains.
A total of 89 samples (74.2%) were reactive for TOSV IgG in at least one of the commercial assays, and 31 samples (31.3%) were reactive in VNT for various SFV serotypes. Average percentage agreements among commercial assays and between VNT and the commercial assays were noted as 57.8% and 62.6%, respectively. No significant correlation between assay results and VNT titers was observed. SFV IgM antibodies were detected in a total of eight samples (6.7%) via IIFT, which were nonreactive in enzyme-linked immunosorbent assay and VNT.
Commercial diagnostic immunoassays displayed slight to fair agreement for TOSV IgG as assessed via kappa and percentage agreement values. The results could only be confirmed via virus neutralization in a portion of the samples, and overall agreement between the commercial assays and VNT was slight. Commercial assays such as immunoblot can be used in addition to VNT for confirmation of TOSV exposure.
[Show abstract][Hide abstract] ABSTRACT: Dengue virus (DENV) and yellow fever virus (YFV) are two of the globally prevalent vector-borne flaviviruses. Data on these viruses from Turkey is limited to a single study originating from the western, Aegean region of Turkey, where evidence for DENV exposure had been confirmed in residents and presence of hemagglutination inhibiting antibodies against YFV had been revealed. The aim of this study was to investigate the rates of seropositivity of DENV and YFV in blood donors from Central/Northern Anatolia, Turkey, for the demonstration of possible human exposure. Serum samples were collected by the Turkish Red Crescent Middle Anatolia Regional Blood Center from donation sites at Ankara, Konya, Eskişehir and Zonguldak provinces and included in the study after informed consent. Ankara is the capital and second most-populated city in Turkey. All samples were previously evaluated for West Nile and tick-borne encephalitis virus antibodies and found to be negative. A total of 2435 and 1502 sera have been evaluated for IgG antibodies against DENV and YFV, respectively. Commercial enzymelinked immunosorbent assays (ELISAs) and indirect immunofluorescence tests (IIFTs) were applied (Euroimmun, Germany) for DENV/YFV IgG surveillance. DENV IgG reactive sera were further evaluated for IgM by ELISA and a commercial mosaic IIFT to determine DENV subtypes. IgM positive samples were also analyzed by a commercial NS1 antigen detection assay (Bio-Rad Laboratories, France). YFV IgG reactive samples were evaluated by IIFT for IgM and via mosaic IIFT and antibody specificity were confirmed by plaque reduction neutralization test (PRNT). Anti-DENV IgGs were demonstrated in repeated assays in 0.9% (21/2435) of the sera. In two samples with borderline IgG results, presence of DENV IgM was detected, one of which was also borderline positive for DENV NS1 antigen. In 14.3% (3/21) of the IgG reactive sera, mosaic IIFT was evaluated as positive and displayed prominent reactivity for DENV-2 in all samples. From five donors with DENV reactivity, new samples were obtained after at least six months which revealed the continuing presence of DENV IgG activity in four. One sample which was initially positive for IgM, borderline for NS1 antigen and borderline for IgG was observed to be positive for IgG and negative for IgM in redonation. IIFT results in three redonation samples also indicated reactivity for DENV-1 and DENV-2 subtypes. Anti-YFV IgGs were detected in 0.6% (9/1502) of the sera. YFV IgM could not be demonstrated in any of the IgG reactive samples and PRNT was evaluated as negative. In conclusion, evidence for DENV exposure, presumably to DENV-2, was identified in residents from Central Anatolian provinces of Ankara and Konya for the first time, however, seroreactivity detected against YFV could not be confirmed by PRNT. These findings indicated that DENV or an antigenically-similar flavivirus was probably present in the study region and sporadic human exposure might have occurred.
[Show abstract][Hide abstract] ABSTRACT: West Nile virus (WNV) is a reemerging flavivirus that has displayed a drastic change in epidemiology in the last decade. Data on WNV activity in Turkey are currently limited. This study investigated WNV exposure in blood donors from Central Anatolia, Turkey.
A total of 2516 sera, collected from blood donors at four major branches of the Turkish Red Crescent Middle Anatolia Regional Blood Center, were evaluated by a commercial WNV immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA). Positive and borderline samples were investigated further by a WNV IgG indirect immunofluorescence test (IIFT), IgG ELISAs for tick-borne encephalitis virus and dengue virus, an IgG IIFT for yellow fever virus, and a multi-Flavivirus biochip IgG IIFT. WNV antibody specificity and titer values were determined by plaque reduction neutralization assay. IgG avidity and IgM were determined for confirmed samples. IgM-positive samples were also evaluated by a real-time reverse transcription polymerase chain reaction assay.
Twenty-five samples (25/2516; 0.99%) were found reactive in the WNV ELISA/IIFT assays, and 14 could be confirmed by the plaque reduction neutralization assay (14/2516; 0.56%). All IgGs were of high avidity, and four samples (4/14; 28.6%), which were negative for viral RNA, were IgM positive. Although samples with neutralizing WNV IgGs had strong fluorescence intensity in IIFTs, no correlation between antibody titer values and IIFT intensity or quantitative ELISA results could be found. Three WNV nonreactive samples were positive in the dengue IgG ELISA test; one of these also displayed positive results for dengue virus in the mosaic biochip IIFT and reactivity in yellow fever virus IIFT.
WNV exposure is confirmed in 0.56% of the tested healthy blood donors in Central Anatolia, with evidence for dengue/yellow fever and/or other flaviviral infections. This study is the first to document WNV exposure in individuals from Konya, Yozgat, and Sivas provinces in Central Anatolia, and it also establishes viral activity in Ankara, the capital of Turkey.