[Comparison of Two Anti-HIV Confirmatory Assays: Recombinant HIV 1/2 Line Immunoassay and Geenius HIV 1/2 Confirmatory Test]. / Iki Anti-HIV Dogrulama Testinin Karsilastirilmasi: Rekombinant HIV 1/2 "Line Immunoassay" ve Geenius HIV 1/2 Dogrulama Testi.

The main purpose in the diagnosis of human immunodeficiency virus (HIV) infection is to rapidly and accurately identify people with HIV infection. It is recommended that samples that are repeatedly reactive should be verified/supported according to the classical algorithms of international and national guidelines. The recombinant "line immunoassay test (LIA)", which has been used for many years, is studied with the accumulated samples to be cost and labor-effective. In this study, the supplemental recombinant HIV 1/2 LIA (INNO-LIA®, Fujirebio, Ghent, Belgium) used to confirm and differentiate the diagnosis of HIV-1 and HIV-2 infections, and an immunochromatographic supplemental test (Geenius™ (Bio-Rad Laboratories, Marnes-la-Coquette, France) which can provide faster results were compared. One hundred fifty serum samples sent to Ege University Faculty of Medicine Hospital Medical Virology Laboratory with anti-HIV 1/2 and p24 antigen positive and indeterminant results and three HIV-1 positive external quality control samples were included in the study. Samples were tested both with the Geenius™ HIV 1/2 (Bio-Rad Laboratories, Marnes-la-Coquette, France) and recombinant HIV 1/2 LIA (INNO-LIA®, Fujirebio, Ghent, Belgium). HIV 1 viral load was evaluated by using Abbott real-time HIV-1 test in Abbott m200sp system (Abbott Molecular, Wiesbaden, Germany) in plasma samples. In both assays, the results were consistent in 147 samples (96.08%). Six samples that have discordant results were as follows: one sample was LIA HIV-1 positive and Geenius indeterminate, two samples were LIA indeterminant and Geenius HIV-1 positive, and in three samples, LIA was indeterminate and Geenius negative. In two EIA reactive samples (2/97, 2.06%) and three EIA negative samples (3/53, 5.66%) LIA results were indeterminant. Geenius test, on the other hand, correctly identified HIV positive and negative samples. The immunochromatographic test could be used in the diagnostic algorithm of HIV infection, due to its short application time, not being labor intensive, its ability to distinguish HIV-1/2, its high sensitivity/specificity compared to LIA, and the compliance with LIA. However, it should be noted that in acute HIV infection, all analytical antibody tests, become reactive later than the fourth generation enzyme immunoassays.

HBsAg, Anti-HCV and Anti-HIV Seroprevalance Among Drug Users: a Retrospective Assessment.

INTRODUCTION: Drug abuse and co-occurring infections are associated with significant morbidity and mortality. In regions with high rates of drug usage, infections like hepatitis B virus (HBV), hepatitis C virus (HCV) and human immunodeficiency virus (HIV) are highly prevalent. Epidemiological studies on HBV, HCV and HIV infections among users of illicit drugs are scarce in Turkey. The primary aim of this cross-sectional and retrospective study was to determine the hepatitis B surface antigen (HBsAg), anti-HCV antibody (anti-HCV) and anti-HIV antibody (anti-HIV) seroprevalences in drug users who applied to a psychiatric outpatient clinic of drug addiction of a university hospital. Secondarily, the assessment of HBsAg, anti-HCV and anti-HIV parameters among intravenous drug users was aimed. METHODS: Serum samples of all patients on probation who applied to a psychiatric outpatient clinic of drug addiction of a university hospital between 2013-2017 and sent to the department of medical microbiology for routine serologic testing were included in the study. The serologic results were obtained retrospectively from laboratory records. For the statistical analysis of the data IBM SPSS 20.0 program was used. RESULTS: Among the studied individuals, the ELISA results demonstrated the existence of HBsAg, anti-HCV and anti-HIV in 94 out of 4357 patients (2.2%), 27 out of 4451 patients (0.6%) and 10 (0.2%) out of 4464 patients, respectively. According to the records, 17 of the patients reported intravenous drug usage. Among this patient group, three patients were found to be anti-HCV positive and one patient was found to be anti-HIV positive. CONCLUSION: In our study, the prevalence of HCV and HIV was increased in patients with intravenous drug usage, whereas in non-intravenous drug users the prevalence is similar to the normal population. In order to plan prevention and harm reduction services for this high-risk population, more national data is needed on HBV, HCV and HIV rates among this group.

The Prevalence of Mixed Genotype Infections in Turkish Patients with Hepatitis C: a Multicentered Assessment.

BACKGROUND: HCV virus infections are one of the major health problems in the world that can cause cirrhosis and liver cancer at a higher rate than other hepatitis data. The aim of this study was to determine the prevalence of mixed infections with different HCV genotypes in Turkey and also to evaluate the current HCV genotype and sub-type distributions by a multicentered assessment. METHODS: The HCV genotype data of 17,578 hepatitis C patients collected from 23 centers from different geographic regions covering all Turkey were collected. The data included information about the HCV genotypes in the last 10 years (between 2007 and 2016), demographic properties of the patients and the methods/systems used to determine the genotypes. RESULTS: Two hundred twenty-eight of the patients (1.3%) had mixed genotype. The most common mixed genotype combination was 1b + 4 (0.83%) followed by 1a + 1b (0.26%). Genotype distribution varies according to geographical regions. However, genotype 1 (82.92%) was the most common genotype in all regions and all years. This was followed by genotype 3 (7.07%) and genotype 4 (5.43%). A variety of methods were used by the centers including sequencing, pyrosequencing, real-time PCR, in-house RFLP, reverse hybridization (LIPA), and hybridization. CONCLUSIONS: Infection with mixed HCV genotypes in Turkey is uncommon. Genotype distribution varies according to geographic regions; the most common genotype 1 is encountered all over the country, while genotypes 3 and 4 are only in some of the centers. Since there is limited information about mixed HCV infection, further investigations are needed to determine the clinical importance of mixed HCV infection.

[Evaluation of viral etiology in central nervous system infections from a university hospital point of view in Izmir based on seven years data]. / Santral sinir sistemi enfeksiyonlarinda viral etiyolojinin Izmir'de bir üniversite hastanesinin yedi yillik verileri üzerinden degerlendirilmesi.

The serious diseases of the central nervous system (CNS); encephalitis and meningitis, have high mortality and morbidity rate especially not diagnosed and treated in time. Nucleic acid testing (NAT) is the tool of choice for viral diagnosis in CNS infections. In this study, viral etiological agents found in cerebrospinal fluid (CSF) samples sent to our university hospital virology laboratory for laboratory diagnosis of CNS infections were retrospectively evaluated and results were compared with other reports from our country. Viral etiological agents found in cerebrospinal fluid (CSF) samples sent to Ege University Faculty of Medicine Department of Medical Microbiology Virology Laboratories for laboratory diagnosis of CNS infection between 01.01.2009-31.12.2015 were evaluated retrospectively. A total of 3778 CSF tests were performed for cell culture of enterovirus (EV) in 487 samples and 3291 tests for nucleic acid testing (NAT) by real time polymerase chain reaction (PCR) in herpes simplex virus 1 (HSV1), herpes simplex virus 2 (HSV2), varicella zoster virus (VZV), Epstein-Barr virus (EBV), cytomegalovirus (CMV), human herpes virus 6 (HHV6) and EV. VZV and EV NAT's were performed during the last one and five years period, respectively. NAT positive results for HSV1, HSV2, CMV, EBV, VZV, HHV6 and EV were 1.80% (24/1333), 0.08% (1/1333), 3.28% (19/580), 4.35% (22/506), 0.46% (1/216), 1.05% (5/478) and 3.37% (6/178), respectively. EV was isolated in 30 (6.20%) of 487 CSF samples by viral culture. Positive samples were mainly from pediatric, neurology and infectious diseases clinics as expected. The number of higher positive results were found in samples sentin december (35.3%), july (12.9%) and november (10.6%). Overall 80% of positive samples belonged to patients over 18 years old. When the results of other studies reported from Turkey are examined, although the positivity rates are generally similar, it is seen that the rates specific to certain factors are higher in selected smaller patient groups like HSV1 and EV. Rapid nucleic acid tests like multiplex PCR and microarray will provide more practical and effective laboratory diagnosis approach in CNS infections, since many more microorganisms may be causative agents.

[Investigation of human herpesvirus-8 seroprevalence in blood donors and HIV-positive patients admitted to Ege University Medical School Hospital, Turkey]. / Ege Üniversitesi Tip Fakültesi Hastanesine basvuran kan donörlerinde ve HIV-pozitif hastalarda insan herpesvirusu-8 seroprevalansinin arastirilmasi.

Human herpesvirus 8 (HHV-8), classified in Herpesviridae family, is the etiological agent of Kaposi's sarcoma (KS), primary effusion lymphoma and multicentric Castleman's disease. In contrast to the other herpesviruses, HHV-8 seroprevalence is low in general populations; however, the higher prevalence observed in individuals with immunodeficiencies such as AIDS poses an increased risk for KS. The global distribution of HHV-8 shows great variations, with the highest seroprevalence seen in Africa. The number of studies on the seroprevalence of HHV-8 in Turkey are limited. The aim of this study was to determine the HHV-8 seroprevalences in healthy blood donors and HIV-positive patients, that will contribute HHV-8 seroepidemiological data in our country. This study was designed as a cross-sectional study. A total of 551 healthy donors (76 female, 475 male; age range: 18-65 years) admitted to Ege University Medical School Hospital, Blood Center for blood donation between December 2013-January 2014, and 173 HIV-positive patients (30 female, 143 male; age range: 18-65 years) admitted to infectious diseases outpatient clinic between October 2013-January 2014, were included in the study. A commercial ELISA method (KSHV/HHV-8 IgG ELISA Kit, Advanced Biotechnologies Inc, USA) was used for the detection of IgG antibodies that were structured against HHV-8 lytic antigens. In the study, 29 (29/551, 5.3%) of blood donors and 44 (44/173, 25.4%) of HIV-positive patients, with a total of 73 (73/724, 10.1%) cases were found as HHV-8 seropositive. The difference between blood donors and HIV-positive patients in terms of HHV-8 seropositivity rates was statistically significant (5.3% versus 25.4%; p< 0.05). In both of the study groups, no statistically significant difference was detected between HHV-8 seropositivity with gender and age. When considering HIV-positive patients, no statistically significant difference was observed between HHV-8 seropositivity with the duration of anti-HIV positivity, CD4(+) T cell count, HIV-RNA status and history of having sexually transmitted disease. As a result, HHV-8 seroprevalence rate detected in our study is similar to the data of other studies performed in Turkey, as well as the rates reported from other European and Asian countries.

[HIV-1 subtype distribution determined by phylogenetic analysis of pol gene sequences and automated subtyping tools among HIV-1 isolates from the Aegian Region of Turkey]. / Ege Bölgesi'nde izole edilen HIV-1 izolatlarinin alttip dagiliminin pol gen bölgesi filogenetik analizi ve otomatize araçlar kullanilarak belirlenmesi.

Human immunodeficiency virus (HIV) exhibiting remarkable genetic variability, includes two genotypes namely HIV-1 (group M, N, O and P) and HIV-2 (group A-H). HIV-1 group M, which is mainly the cause of the AIDS pandemic, is divided into nine pure subtypes, more than 45 circulating recombinant forms (CRF) and numerous unique recombinant forms (URF). According to the documents of Turkish Government of Health, among a total of 6802 HIV-positive cases, 1096 of them were defined as AIDS as of June 2013 in Turkey. Although subtype B is the predominant subtype, recent studies indicate higher proportion of CRFs similar to their increasing role in the HIV pandemic. The aim of this study was to determine the subtype distribution of HIV-1 strains isolated from 70 patients (61 male, 9 female; age range: 16-73 yrs, mean age: 39.6 yrs) who presented to our institution between April 2008-June 2013. HIV-1 strains were subtyped by phylogenetic analysis of the pol gene region and commonly used automated subtyping tools namely, Stanford HIV db v6.2.0 and Rega v3.0. Pol sequences retrieved from the Los Alamos database and from GeneBank, were trimmed from full-length genomes. Phylogenetic analysis of the 1302 base pair of the pol gene region was performed using Mega v5.2 software. The sequences were aligned using Muscle and phylogenetic distances between sequences were estimated by using Kimura two-parameter model (transition/transversion ratio: 2.0). Tree topology was obtained using neighbour-joining method and bootstrap value was set at 1000. Sixty-one (87.1%) patients were antiretroviral treatment (ART)-naive and nine were on different ART regimens. The subtypes of the isolates according to phylogenetic analysis were found as follows; 31 (44.2%) subtype B, 24 (34.2%) CRF42_BF, 6 (8.5%) B/CRF02_AG recombinants, 5 (7.1%) sub-subtype A1, 1 (1.4%) sub-subtype F1, 1 (%1.4) CRF 25_cpx, 1 (1.4%) CRF02_AG and 1 (1.4%) CRF01_AE. Rega v3.0 subtyping tool produced five discrepant results (4 B/CRF02-AG and 1 CRF42_BF) compared to phylogenetic analysis. Stanford HIVdb v6.2.0 had eight results (3 CRF42_BF, 2 subtype B, 2 sub-subtype A1, 1 CRF25_cpx) that were not concordant with phylogenetic analysis. Stanford HIVdb v6.2.0 was able to subtype all B/CRF02_AG recombinant strains. B/CRF02_AG recombinants which were seen among homosexual men in France were for the first time isolated in Turkey from five men (2 homosexual, 2 bisexual, 1 heterosexual) and one heterosexual woman. CRF42_BF had not been found in Turkey previously and it has not been a common type isolated in neighboring countries either. Full genome sequencing could be helpful to further analysis of those isolates. Our results support the latest studies from Turkey reporting increase in the proportion of CRF-related infections. This is not an unusual finding when geographical location of Turkey is considered. Nevertheless, more comprehensive data regarding molecular epidemiology and subtype distribution of HIV-1 isolates in Turkey are needed.

[Investigation of hepatitis C virus genotype distribution in patients with chronic hepatitis C infections in Antalya Training and Research Hospital, Turkey]. / Antalya Egitim ve Arastirma Hastanesinde kronik hepatit C hastalarinin genotip dagiliminin arastirilmasi

Hepatitis C virus (HCV) infection is a major global health problem due to high chronicity rates, occurrence of severe hepatic diseases, and absence of an accurate therapy and effective vaccine. It is well known that viral genome is highly variable and HCV has at least six genotypes, each of them containing a series of subtypes. HCV genotypes exhibit geographical and epidemiological distribution. Genotype identification is clinically important to decide the dosage and duration of treatment since different genotypes exhibit variable response to treatment. The aim of this study was to determine the HCV genotypes in chronic HCV patients who were followed-up in Antalya Research and Training Hospital, Turkey. Anti-HCV and HCV-RNA positive blood samples obtained from 148 chronic hepatitis C patients (67 female, 81 male; mean age: 50.5 ± 10.8, age range: 17-73 years) who were admitted to Antalya Research and Training Hospital Microbiology Laboratory during January 2011-June 2013, were included in the study. Epidemiological data of the patients and HCV genotype results were evaluated retrospectively. Viral genotypes were determined by real-time (Rt) PCR assay (Abbott Molecular Diagnostic, USA). HCV genotype (Gt)-1 was detected in 119 (80.4%) of the patients, of them 15.9% (19/119) were identified as subtype 1a and 75.6% (90/119) were subtype 1b. The prevalence rates of Gt-2, -3, and -4 were found as 3.4% (n= 5), 11.5% (n= 17), and 2% (n= 3), respectively. Gt-6 was not detected in our patients. Mixed infection with HCV types was detected in four patients (2.7%) by Rt-PCR; of these three were detected as Gt-1 and one was Gt-2 by RFLP (Restriction Fragment Length Polymorphism) and sequencing. The high prevalence of Gt-3 (11.5%) obtained in this study was attributed to the determination of Gt-3 in seven of 13 foreign national subjects. Rt-PCR method used in this study is user independent, standardized, automated, rapid and reliable method, however in case of detection of mixed types, the samples should be confirmed by other methods. In conclusion, we reported that the majority of the chronic hepatitis C infected patients had Gt-1b, and Gt-3 exhibited the highest rate ever reported by other studies from Turkey.

Possible transmission risks and genotype distribution of hepatitis C virus infection in Western Turkey.

BACKGROUND/AIMS: The aim of this study was to investigate the risk factors which may be involved in the transmission of hepatitis C virus and to determine the recent distribution of various genotypes in Western Turkey. MATERIALS AND METHODS: The risk determination study consisted of 215 patients whose serum samples were sent to the Medical Microbiology Laboratory at Ege University Hospital between 2005 and 2010 and were anti-hepatitis C virus positive. For the determination of recent genotype distribution, genotyping results of all 535 patients sent to the same laboratory from 2007 to 2011 were analyzed. Information on possible risk factors for the transmission of hepatitis C virus was obtained by a telephone questionnaire. Hepatitis C virus typing was performed by restriction fragment length polymorphism analysis. RESULTS: The most frequently reported risk factors were history of dental procedures in 171 (79,5%) patients and surgical operations in 137 (63,7%) patients. Genotype 1 was observed in 499 of the 535 patients (93,3%) with chronic hepatitis C virus infection. Of these, 69 patients showed infection with subtype 1a (12.9.%) and 430 - with subtype 1b (80.4%). Genotype 3 was determined in 20 patients (3,7%), genotype 2 - in 8 patients (1,5), and genotype 4 - in 8 patients (1,5%). CONCLUSIONS: Even though there is an increase in non-1 genotypes, Turkish patients with chronic hepatitis C still represent a rather homogenous group with genotypic diversity encountered rarely. The risk factors detected in the patients admitted to our hospital are mainly medical procedures which can be prevented by the use of simple infection control practices and implementation of an education program.

[Role of line immunoassay in the diagnosis of early HIV infection: a diagnostic case]. / Erken dönem HIV enfeksiyonunda "line immunoassay" testinin yeri: Tanisal bir olgu.

Combined p24 antigen-HIV antibody fourth-generation assays that identify most of the early HIV infections have been used extensively worldwide for several years. This poses challenges for the traditional algorithm of line immunoassay (LIA) confirmation. LIA tests are useful methods with their high specificity and their ability to differentiate HIV-1 from HIV-2, but they are reactive days after the fourth generation enzyme immunoassays. With acute HIV infection, high levels of infectious virus are detectable in serum and genital secretions. The rate of transmission during acute HIV infection is higher than the established HIV infection, for this reason, new HIV testing strategies need to focus on sensitivity, especially for this highly contagious phase immediately after infection. Serum sample of a patient sent to Ege University Hospital Clinical Virology Laboratory was repeatedly reactive with low signal/cutoff ratios with two different commercial fourth generation enzyme immunoassays (Architect HIV Ag/Ab Combo Reagent Kit, Abbott, Germany and Vidas HIV Duo Quick, Biomerieux, France). The sample was non-reactive with the LIA (INNO-LIA HIV I/II Score, Innogenetics, Belgium) and HIV RNA (RealTime HIV-I Amplification Reagent Kit, Abbott, USA) result was positive (4.1 x 10(5) copies/ml). With the presentation of this case, the role of LIA in the diagnosis of early HIV infection and its place in test algorithms were questioned.

[Subacute sclerosing panencephalitis cases diagnosed by increased CSF/serum measles antibody indices]. / Yüksek BOS/Serum Kizamik Antikor Indeksi ile Tanimlanan Subakut Sklerozan Panensefalit Olgulari

Subacute sclerosing panencephalitis (SSPE) caused by persistent defective measles virus strains, is a progressive neurological disorder of children and adolescents. The aim of this letter was to share the data from SSPE-suspected cases who were definitely diagnosed by the detection of increased antibody index in serum and cerebrospinal fluid (CSF) samples. A total of 11 patients (mean age: 14.3 years) with suspected SSPE between February 2006 to August 2008, were included in the study. Simultaneously obtained serum and CSF samples from patients were analyzed in terms of albumin, total IgG and measles-specific IgG levels (Measles Virus IgG ELISA for CSF Diagnostics, Euroimmun, Germany). The value of CSQrel (relative CSF/serum quotient) ≥ 1.5 was accepted indicative for intrathecal measles antibody synthesis. Seven (63.6%) of the 11 patients' diagnosis were confirmed with the demonstration of elevated CSF/serum indices (CSQrel range: 2.3-36.9; mean: 12.9). Mean age of those seven cases was 12.3 years (age range: 7-21) and four of them were male. The history of patients with high antibody indices indicated that three of four patients who had measles infection had not been vaccinated against measles. These three unvaccinated patients had measles infection at 3rd, 8th and 30th months of age, respectively, and the period of SSPE development were 15, 6 and 4.5 years, respectively. With this letter we would like to emphasize once more that effective measles vaccination is the only way for the prevention of measles and SSPE and the demonstration of increased measles antibody index in simultaneously obtained serum and CSF samples is crucial for the diagnosis of SSPE.

[Identification of enteroviruses from central nervous system infections by RT-PCR and cell culture methods]. / Santral sinir sistemi enfeksiyonu etkeni enteroviruslarin RT-PCR ve hücre kültür yöntemleri ile saptanmasi.

Viruses are the major causes of aseptic meningitis and encephalitis. Enteroviruses account for more than 80% of the aseptic meningitis cases for which an etiologic agent is identified. The aims of the present study were to identify agents of enteroviral meningitis by viral culture and reverse transcriptase polymerase chain reaction (RT-PCR) methods, to evaluate the appropriateness of a commercial RTPCR kit for its use in routine laboratory, and to obtain epidemiological data about enteroviral meningitis. Sixty six cerebrospinal fluid (CSF) samples from patients with suspected viral central nervous system (CNS) infection by clinical and CSF biochemical findings, sent to Ege University Faculty of Medicine, Department of Medical Microbiology were included in the study. The CSF samples were all negative for tested bacteria, mycobacteria, fungi, herpes simplex virus and cytomegalovirus. Thirty-four (51.5%) of the samples were from female and 32 (48.5%) were from male patients. Twenty-three (34.8%) patients were children (5 months-18 years) and 43 (65.2%) were adults (19-86 years). Shell vial rapid cell culture method by using Vero, HEp-2 and RD cell lines was performed for virus isolation and the results were evaluated on 48th hours after staining the cells with fluorescein labeled polyclonal antibodies (Pan-Enterovirus Blend, Light Diagnostics, USA). Enteroviral RNA in the samples was detected by a commercial RT-PCR kit (Enterovirus Consensus Kit, Argene, France). Sixty-one (92.4%) of 66 samples from patients with suspected viral CNS infection were found to be negative for enterovirus both with RT-PCR and shell vial cell culture methods. Three samples (4.5%) were positive by shell vial culture method. In one CSF sample that was culture positive, RT-PCR was also positive. However, the remaining two culture positive samples yielded negative result by RT-PCR. Intermediate results with RT-PCR were obtained in two samples (3%) that were identified as negative by cell culture. Two of the three positive samples in cell culture were identified as echovirus, however, the remaining sample could not be identified due to small sample amount. As a result, the commercial assay was found non-practical and labor intensive, giving indeterminant results in some cases and missing two culture positive samples. Since it didn't have an advantage over the cell culture method used, it was found inappropriate for routine diagnosis in our laboratory. On the other hand, it has been known that nucleic acid amplification tests (NAT) have markedly improved the diagnosis of enterovirus infections by increasing the sensitivity compared with cell culture methods. An alternative NAT method should be evaluated in parallel with cell culture method especially in CSF samples of children with suspected viral central nervous system infections.

[Evaluation of immunoblot-based assay for detecting Epstein-Barr virus viral capsid antibodies]. / Epstein-Barr virus viral kapsid antikorlarinin saptanmasinda immünoblot yönteminin degerlendirilmesi.

Various attempts have been made to improve Epstein-Barr virus (EBV) serodiagnosis by developing more practical and objective methods than immunofluorescence-based assays. In the present study, the performance of immunoblot-based assays were evaluated by comparing the results obtained by the gold standard immunofluorescence antibody (IFA) test for the detection of IgM and IgG antibodies against EBV viral capsid antigen (anti-VCA). Serum samples of 277 patients admitted to Ege University Hospital for routine EBV diagnosis were included in the study. The age range of the patients was 3 months-89 years (mean 28 years) and 104 of them were females and 173 were males. All the samples were assayed by commercial immunoblot (Euroline IgM and IgG; Euroimmun, Germany) and IFA (EBV-CA IgG and IgM, Euroimmun, Germany) methods. Crosstabulation, chi-square test and phi (phi) measures in SPSS 16.0 statistical package programme were used for data analysis. Of the 216 samples that were interpreted as positive with immunoblot-based IgM assay, only 34 (15.7%) were confirmed as positive with IFA, whereas 162 (75%) were negative, and 20 (9.3%) were equivocal (phi = 0.167; low correlation). Of the 85 samples that were anti-VCA IgG positive with immunoblot assay, 82 (96.5%) were positive, 2 (2.3%) were negative and 1 (1.2%) were equivocal with IFA (phi = 0.441; significant correlation). When the indeterminate results obtained by IFA test were excluded from the evaluation, the correlation between immunoblot VCA IgG and IFA IgG was 85.4% (88/103) and between immunoblot VCA IgM and IFA IgM was 27.3% (69/253). When the intensities of bands were evaluated for IgM testing, it was noted that as the intensity of the bands increased (1+ to 3+), IFA VCA IgM reactivity rates increased (from 9.9% to 29.5% for p19 band; from 24% to 85.7% for gp125 band). For immunoblot VCA IgM testing, 165 samples were found to be positive only for VCA p19 band. Of these samples, 135 (81.8%) were negative, 15 (9.1%) were positive and 15 (9.1%) were equivocal with IFA. It is observed that even though immunoblot assays with automated blotting and scanning systems can be a convenient alternative to immunofluorescence assay, the rate of false positivity obtained for VCA IgM was high (75%). It was concluded that in laboratories which apply immunoblotting as a primary screening test for EBV serodiagnosis, the positive VCA IgM results (particularly isolated p19 band positivity) and the presence of low intensity bands, should be confirmed by IFA testing.

Comparison of immunofluorescence assay and multiplexed microparticle-based immunoassay for detecting Epstein-Barr virus viral capsid antigen antibodies.

A new multiplexed microparticle-based immunoassay was compared with the immunofluorescence assay that is used widely for detecting EBV-specific antibodies in immunocompetent patients. Serum samples of 162 patients submitted for routine EBV diagnosis were tested for viral capsid antigen IgM, viral capsid antigen IgG and serological profile interpretations with both systems. The result concordances were 94.2%, 93.6%, and 92.1%, respectively. Multiplexed microparticle-based immunoassay can be an alternative to immunofluorescence assay especially in laboratories receiving large numbers of samples.

Distribution of hepatitis C virus genotypes in patients with chronic hepatitis C infection in Western Turkey.

OBJECTIVE: The primary aim of this study was to determine the recent distribution of various genotypes of hepatitis C virus (HCV) in patients with chronic HCV infection in Western Turkey. Additional objectives were to determine whether there are any associations of genotype with gender and age, and to determine the nucleotide similarities and risk factors of non-1 HCV genotypes. METHODS: Serum samples from 345 patients (176 male, 169 female; mean age 53.3+/-12.7 years, range 10-81 years) with chronic HCV infection were analyzed in this study. Viral genotypes were determined by a restriction fragment length polymorphism (RFLP)-based in-house assay. To confirm genotypes for the samples with band patterns other than genotype 1, the 5' UTR was amplified and sequenced. RESULTS: Genotype 1 was observed in 335 of the 345 patients (97.1%). Of these, 34 patients showed infection with subtype 1a (9.9%) and 301 with subtype 1b (87.2%). Genotypes 2, 3, and 4 were determined in 0.9%, 1.4%, and 0.6% of the patients, respectively. Patients infected with type 1 were significantly older than patients infected with non-1 genotypes; however no significant differences were recorded in gender distribution. CONCLUSIONS: Genotypes other than genotype 1 are quite rare; these are possibly acquired in other countries. Turkish patients with chronic hepatitis C still represent a rather homogenous group with genotypic diversity encountered rarely.

Efficacy of accelerated hepatitis B vaccination program in patients being actively treated for hematologic malignancies.

BACKGROUND: The goal of this study was to conduct an accelerated vaccination program and to determine its efficacy in patients susceptible to hepatitis B virus (HBV) receiving chemotherapy because of their hematologic malignancies. METHODS: Over a one-year period, a total of 327 patients who were diagnosed as having a hematologic malignancy were serologically analyzed in terms of HBV infection. Of those found to be susceptible to HBV infection, a total of 42 patients consisting of 16 females and 26 males were enrolled in the accelerated vaccination program. All the patients were administered a 20-microg yeast-derived recombinant hepatitis B vaccine on days 0, 14, and 28. Anti-HBs titers above 10IU/l at 1 and 3 months after the final dose were accepted as protective. RESULTS: A total of 146 (44.6%) patients were susceptible to HBV, while 13 (4.0%) were carriers, 28 (8.6%) were vaccinated, and 113 (34.5%) had had a previous HBV infection. A total of 42 patients (16 females and 26 males, mean age 34.5+/-10.9 years) were enrolled in the vaccination program. Overall, 23.8% (10/42) of the patients in the program had developed anti-HBs at one month after the last vaccination. CONCLUSIONS: Poor results obtained by different vaccination programs suggest the need for alternative strategies to prevent the disease.

Evaluation of three different assays for the assessment of Epstein Barr Virus immunological status.

Various attempts have been made to improve Epstein Barr Virus serodiagnosis by developing convenient methods. The present study evaluated the performance of multiplexed bead assays and immunoblot based assays on automated platforms by comparing them with immunofluorescence based assays for the determination of EBV immune status. A total of 45 serum samples were included in the study. Serum samples were tested by multiplexed bead EBV assays (AtheNA Multi-Lyte, Zeus Scientific,USA) and immunoblot based assays (Euroline, Euroimmun AG, Germany) on automated platforms. Assay systems were evaluated by comparing them with immunofluorescence based assays (Zeus Scientific, USA). For EBV anti-VCA IgM, anti-VCA IgG, anti-EA and anti-EBNA, the kappa values reflecting agreements of AtheNA and IFA were 0.20, 0.54, 0.92 and 0.95 for anti-EA, anti-VCA IgG, anti-VCA IgM and anti-EBNA respectively and the agreements of Euroline and IFA were 0.53, 0.67, 0.81 and 1.000 for anti-VCA IgG, anti-EA, anti-VCA IgM and anti-EBNA respectively. The results of the study performed on a limited number of serum samples demonstrated that the multiplexed bead assays and immunoblot assays agree with the standard IFA assay for anti-EBNA IgG and anti-VCA IgM detection while the agreement is less for anti-EA and anti-VCA IgG.

Comparison of ELISA with shell vial cell culture method for the detection of human rotavirus in fecal specimens.

The aim of the study was to compare an enzyme immunoassay method with shell vial cell culture method for detection of rotavirus in fecal specimens. In addition, the correlation between laboratory results and clinical scores of patients with gastroenteritis was evaluated. A total of 219 fecal specimens from children (ages 3 weeks to 5 years) with acute gastroenteritis submitted to pediatric emergency room were evaluated by both ELISA and shell vial cell culture. A Vesikari score was used for assessing the severity of the illness. Among 219 stool samples tested, 107 (48.9%) were determined to be positive. Two specimens were positive by shell vial cell culture method while they were ELISA negative. According to these results the calculated sensitivity, specificity, PPV, and NPV of ELISA were 98.1%, 100%, 100%, and 98.2%, respectively. The mean severity score for the 107 episodes of rotavirus diarrhoea was 11.0 +/- 3.6 compared to 4.5 +/- 1.9 for the 112 episodes of non-rotavirus diarrhea in the same population. Our study indicates that ELISA, which is easier to perform, faster and cheaper than cell culture methods may be suitable for routine diagnosis of rotavirus infections. The severity of rotavirus positive gastroenteritis was significantly higher than that of rotavirus negative patients.

[Development of a database for tracking HIV positive/AIDS patients]. / HIV pozitif/AIDS hastalarinin tani ve izlemi için gelistirilen veri tabani ortami.

The collection of reliable data is the first step to assess the status of HIV/AIDS in a community. HIV recording systems are necessary for organizing and analyzing the patients' data. The aim of the study was to develop a database to be used to track HIV positive/AIDS patients. The database includes general demographic fields as well as specific fields such as health history, laboratory and other clinical history, current and past drug regimens (both antiretroviral and non-antiretroviral drugs). It is also possible to organize and maintain a patient database according to specific diseases, laboratory tests and/or medication treatments.

Hepatitis delta virus (HDV) genotypes in patients with chronic hepatitis: molecular epidemiology of HDV in Turkey.

OBJECTIVE: Analysis of hepatitis delta virus (HDV) isolates from around the world has indicated that there are at least three phylogenetically distinct genotypes with different geographic distributions. The aim of this study was to determine the distribution of HDV genotypes by direct sequencing in patients with chronic delta hepatitis in Izmir, Turkey. DESIGN AND METHODS: Serum samples from 32 chronic hepatitis patients (21 males, 11 females; mean age 44.2 years, range 23-70 years) with anti-delta positivity were analyzed for hepatitis B and C serologies. After reverse transcription, cDNA of partial delta antigen was amplified by in-house nested PCR. The products of the HDV PCR were bidirectionally sequenced with internal primers using Big Dye Terminator DNA Sequencing Kit (Applied Biosystems, CA, USA) and ABI Prism 310 Genetic Analyzer (Perkin Elmer, USA). Nucleotide sequences of HDV were compared with previously reported sequences and aligned by using ClustalW (1.82). RESULTS: HDV-RNA was positive in 26 (81.3%) of 32 anti-delta positive samples. Comparison of the HDV sequences with published sequences of HDV genotypes I, II, and III indicated that all were closely related to HDV genotype I isolates. Similarity among isolated sequences ranged from 84% to 96%. CONCLUSION: HDV genotyping was successfully performed by direct sequencing of the amplicons obtained from routine HDV-RNA screening PCR tests. All of the HDV isolates from the chronic delta hepatitis patients included in this study were found to be genotype I.