West Nile virus seropositivity in Alanya, a coastal city in the Mediterranean region of Turkey.

BACKGROUND: West Nile virus (WNV)-related illness is a global health problem. Understanding the seropositivity rates and identifying the risk factors related to WNV in various animal species including humans is crucial for the implementation of effective prevention strategies. OBJECTIVES: Assess the rate of seropositivity and the risk factors associated with WNV seropositivity. DESIGN: Descriptive, cross-sectional. SETTING: Microbiology and virology departments in a veterinary college. PATIENTS AND METHODS: In a sample of healthy human participants in Alanya, located close to regions where WNV activity has been detected, anti-WNV IgG antibody detection was performed using enzyme-linked immunosorbent assays. The positive results were confirmed by virus neutralization tests (VNTs). The sample was compared with a second group of age- and gender-matched healthy subjects selected from a previous cross-sectional study. MAIN OUTCOME MEASURES: Determination of the seropositivity and risk factors that were associated with WNV in healthy humans. SAMPLE SIZE: 87 in current study; 356 in previous study. RESULTS: The first group of 87, which had a high risk of encountering vector mosquitoes, had a positivity rate of 8% (7/87), whereas positivity in the second group was 4.5% (16/356; P=.181). In the entire sample, the anti-WNV IgG antibody was positive in 23 out of 443 (5.2%) samples by the ELISA test. Among these 23 samples, ten were confirmed as positive using VNTs. Therefore, the WNV IgG seropositivity was 2.3% (10/442). Confirmed IgG seropositivity rates were higher among male (3.8%) than female participants (0.9%; P=.054) and among adults aged ≥45 years (4%) than those aged 18-44 years (0.8%; P=.048). CONCLUSION: This study highlights the presence of WNV infection in the research region. More comprehensive and multidisciplinary studies are required to increase our knowledge about this zoonotic infection including risk factors in line with the One Health approach. LIMITATIONS: Small sample size.

[Viral Respiratory Tract Infection Agents Detected Between Years 2019-2021, COVID-19, and Co-Infections]. / 2019-2021 Yillari Arasinda Saptanan Viral Solunum Yolu Enfeksiyonu Etkenleri, COVID-19 ve Ko-Enfeksiyonlar.

Respiratory tract infections are a major cause of morbidity and mortality at all ages and are seen as a very important public health problem all over the world. In this study, we aimed to evaluate the effect of the pandemic on the epidemiological and seasonal characteristics of the agents by analyzing the respiratory viral infection agents, viral co-infections and associations with Coronavirus diseases-2019 (COVID-19) studied by multiplex polymerase chain reaction (PCR) test in the molecular microbiology laboratory in a three-year period, including the one-year period before the pandemic. Between March 2019 and December 2021, 8825 respiratory tract specimens accepted to the molecular microbiology laboratory with respiratory tract multiplex PCR test requests were included in the study. In addition, severe acute respiratory syndrome (SARS-CoV-2) PCR test results of the patients with positive results with respiratory tract multiplex PCR test, which were studied within ± 3 days, were evaluated retrospectively. Respiratory viral pathogens were detected using FTD Respiratory Pathogens 21 kit (Fast Tract Diagnostics, Siemens Healthineers Company). Two different kits based on real-time reverse transcription PCR were used for SARS-CoV-2 RNA detection in different periods. According to our results, at least one viral agent was detected in 2156 (24.4%) of a total of 8825 samples and a single agent was detected in 1843 (85.5%) of these. The distribution of viruses in the samples with a single agent was determined as RV, RSV A/B, HCoVs, AdV, flu A virus, MPV A/B, PIV 1-4, flu B virus, EV, BoV and PeV, in order of frequency. Multiple agents were found in 313 (14.5%) of these 2156 samples. They were found to be two agents in 291 samples, three in 21 samples and four in one sample. When the SARS-CoV-2 PCR test results of the patients who had positive results with respiratory tract multiplex PCR and who were studied within ± 3 days were evaluated retrospectively, SARS-CoV-2 RNA was detected in 45 (3.5%) of 1277 samples in which at least one agent was detected. In four of these patients, SARS-CoV-2 was found together with multiple agents. Consequently, there was a sharp decrease in the prevalence of all viral agents during the pandemic period. It was evaluated that besides the COVID-19 infection, the restrictions applied during the pandemic period were also effective in this situation.

Impact of non-pharmaceutical interventions on circulating respiratory viruses during the COVID-19 pandemic in Turkey.

BACKGROUND: Non-pharmaceutical interventions (NPIs) applied to limit the SARS-CoV-2 pandemic also affect the circulation and seasonal characteristics of other respiratory viruses. OBJECTIVES: Assess the impact of NPIs on the spread and seasonal characteristics of non-SARS-CoV-2 respiratory viruses and examine viral respiratory co-infections. DESIGN: Retrospective cohort SETTING: Single center in Turkey. PATIENTS AND METHODS: Syndromic multiplex viral polymerase chain reaction (mPCR) panel results of patients admitted to the Ankara Bilkent City Hospital with symptoms of acute respiratory tract infection between April 1, 2020 and October 30, 2022 were evaluated. Two study periods before and after 1 July 2021, when the restrictions were discontinued, were statistically analyzed and compared to determine the effect of NPIs on circulating respiratory viruses. MAIN OUTCOME MEASURES: Prevalence of respiratory viruses as determined by syndromic mPCR panel. SAMPLE SIZE: 11300 patient samples were evaluated. RESULTS: At least one respiratory tract virus was detected in 6250 (55.3%) patients. Of these, at least one respiratory virus was detected in 5% in the first period (between April 1, 2020 and June 30, 2021, when NPIs were applied), and in 95% in the second period (between July 1, 2021 and October 30, 2022, when NPIs were relaxed). After the removal of NPIs, there was a statistically significant increase in hRV/EV, RSV-A/B, Flu A/H3, hBoV, hMPV, PIV-1, PIV-4, hCoV-OC43, PIV-2 and hCoV-NL63 (P<.05). In the 2020-2021 season, when strict NPIs were applied, all respiratory viruses evaluated did not have the usual seasonal peak and there were no seasonal influenza epidemics during this period. CONCLUSIONS: NPIs resulted in a dramatic decrease in the prevalence of respiratory viruses and notable disruption of seasonal characteristics. LIMITATIONS: Single-center study and retrospective. CONFLICT OF INTEREST: None.

One-Year Post-Vaccination Longitudinal Follow-Up of Quantitative SARS-CoV-2 Anti-Spike Total Antibodies in Health Care Professionals and Evaluation of Correlation with Surrogate Neutralization Test.

Numerous vaccines have been generated to decrease the morbidity and mortality of COVID-19. This study aims to evaluate the immunogenicity of the heterologous boosts by BioNTech against homologous boosts by CoronaVac at three-month intervals in two health care worker (HCW) cohorts, with or without prior COVID-19, for one year post-vaccination. This is a prospective cohort study in which the humoral responses of 386 HCWs were followed-up longitudinally in six main groups according to their previous COVID-19 exposure and vaccination status. Anti-SARS-CoV-2 spike-RBD total antibody levels were measured and SARS-CoV-2 neutralization antibody (NAbs) responses against the ancestral Wuhan and the Omicron variant were evaluated comparatively using international standard serum for Wuhan and Omicron, as well as with the aid of a conversion tool. The anti-SARS-CoV-2 spike-RBD total Ab and Nab difference between with and without prior COVID-19, three months after two-dose primary vaccination with CoronaVac, was statistically significant (p = 0.001). In the subsequent follow-ups, this difference was not observed between the groups. Those previously infected (PI) and non-previously infected (NPI) groups receiving BioNTech as the third dose had higher anti-SARS-CoV-2 spike total Ab levels (14.2-fold and 17.4-fold, respectively, p = 0.001) and Nab responses (against Wuhan and Omicron) than those receiving CoronaVac. Ab responses after booster vaccination decreased significantly in all groups at the ninth-month follow-up (p < 0.05); however, Abs were still higher in all booster received groups than that in the primary vaccination. Abs were above the protective level at the twelfth-month measurement in the entire of the second BioNTech received group as the fourth dose of vaccination. In the one-year follow-up period, the increased incidence of COVID-19 in the groups vaccinated with two or three doses of CoronaVac compared with the groups vaccinated with BioNTech as a booster suggested that continuing the heterologous CoronaVac/BioNTech vaccination, revised according to current SARS-CoV-2 variants and with at least a six-month interval booster would be an effective and safe strategy for protection against COVID-19, particularly in health care workers.

Comparison of clinical characteristics of wild-type SARS-CoV-2 and Omicron.

OBJECTIVE: This study aimed to investigate the effect of mutations by comparing wild-type SARS-CoV-2 and Omicron regarding clinical features in patients with COVID-19. It also aimed to assess whether SARS-CoV-2 cycle threshold value could predict COVID-19 severity. METHODS: A total of 960 wild-type and 411 Omicron variant patients with positive results in SARS-CoV-2 real-time reverse transcriptase polymerase chain reaction test from oropharyngeal and/or nasopharyngeal samples during their hospital admissions were included in this retrospective study. The reference symptoms of the patients were obtained from the hospital database. The correlation between chest computed tomography findings and the "cycle threshold" of patients with wild-type SARS-CoV-2 was assessed. RESULTS: Cough, fever, shortness of breath, loss of taste and smell, and diarrhea were found to be statistically significantly higher (p=0.001; 0.001; 0.001; 0.001; and 0.006; respectively) in the wild-type cohort, while in the Omicron cohort, sore throat and headache were found to be statistically significantly higher (p=0.001 and 0.003, respectively). An inverse relationship was found between chest computed tomography findings and viral load. CONCLUSION: This study revealed that the Omicron variant tended to infect predominantly the upper respiratory tract and showed decreased lung infectivity, and the disease progressed with a milder clinical course. Therefore, the study showed that the tropism of the virus was changed and the viral phenotype was affected. It was also found that SARS-CoV-2 viral load did not predict COVID-19 severity in patients with wild-type SARS-CoV-2.

Sensitivity Affected by Disease Severity and Serum Sampling Time: a Performance Evaluation of Six SARS-CoV-2 Antibody Immunoassays.

Comparative validation and clinical performance data are essential for the reliable interpretation of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) antibody test results. This study aimed to assess the performance of six SARS-CoV-2 IgG immunoassays in the context of different disease severities. Four automated chemiluminescence immunoassays (Access [Beckman Coulter], Architect [Abbott], Atellica-IM [Siemens], and Elecsys [Roche]) as well as two ELISA assays (SARS-CoV-2 IgG-S1-based and NCP IgG [Euroimmun]) were evaluated using samples from 143 patients as well as 50 pre-pandemic control serum samples. Accuracy and precision tests were performed for validation purposes. Overall sensitivity ranged between 73.38-88.65% and was higher in spike protein-based assays, while the specificity was ≥98% in all immunoassays. The clinical performance of the immunoassays differed depending on disease severity and target antigen. For instance, the IgG response was lower for samples taken <20 days post-symptom onset (87.30%) compared with those taken ≥20 days post-symptom onset (94.80%). Moreover, moderate disease levels led to the highest levels of IgG. Higher levels of antibodies were detected in the clinically moderate disease group. In asymptomatic and mild groups, more antibody positivity was detected with spike protein-based assays. All the assays tested could be used to detect SARS-CoV-2 IgG. However, spike-based assays revealed relatively higher sensitivity rates than nucleoprotein-based assays, particularly in cases of asymptomatic and mild disease.

Evaluation of antibody response after COVID-19 vaccination of healthcare workers.

The common goal of all vaccines developed against COVID-19, although they have been designed with different methods, is to develop an effective immunity and antibody response against SARS-CoV-2. However, the postvaccination immune response and antibody levels differ between individuals. This study examined the relationship between postvaccine seropositivity rates, age, gender, smoking, and body mass index (BMI), and antibody titers. A total of 314 healthcare workers (HCW) who were not previously infected with COVID-19 and who had received two doses of CoronaVac inactivated vaccine participated in the study. Seropositivity against the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein was measured from the participants 4 weeks after the second dose of vaccine using the electrochemiluminescence (ECLIA) method. In addition, the antibody developed against the nucleocapsid protein (NCP) was evaluated and compared using Elecsys Anti-SARS-CoV-2 kit. One hundred and eighty-one of the participants were female (57.6%) with a median age of 39 (interquartile range [IQR], 10) and 133 (42.4%) were male with a median age of 41 (IQR, 11). 99.6% of the volunteers developed seropositivity 4 weeks after the second dose of vaccine. It was also observed that the rate of RBD protein antibody titer was >250 U/ml in smokers, which is quite low compared to nonsmokers (p = 0.032), and that high RBD antibody titers were proportionally lower in obese participants, according to BMI values, compared to those with normal BMI (49.5% and 9.9%). It was observed that seropositivity developed in almost all participants after the CoronaVac vaccine. However, it was determined that the antibody titer measured varied depending on factors such as smoking, BMI, and duration.

Molecular Characterization of Carbapenem-Resistant Acinetobacter baumannii Blood Culture Isolates from Three Hospitals in Turkey.

We aimed to investigate the clonal relationships, common sequence types, and carbapenemase genes in 177 non-repetitive blood culture isolates of Acinetobacter baumannii collected from patients at three university hospitals in Turkey in 2016. Molecular epidemiological characteristics of the isolates were examined using pulsed-field gel electrophoresis (PFGE), and multilocus sequence typing (MLST) (Pasteur scheme-cpn60, fusA, gltA, pyrG, recA, rplB, and rpoB). Multiplex PCR was used to investigate the carbapenemase genes, including blaOXA-23-like, blaOXA-24-like, blaOXA-48-like, blaOXA-58-like, blaIMP, blaVIM, and blaNDM. PFGE genotyping yielded 92 pulsotypes with a clustering ratio of 69.7%. As per a ≥85% similarity coefficient, 159 (90.9%) isolates were found to be clonally related. The blaOXA-23-like and blaOXA-58-like genes were identified in 100% and 28.2% of the isolates, respectively. The blaNDM gene was identified in two isolates. The MLST analysis included 54 isolates with different pulsotypes, and 29 sequence types (STs). Most of the isolates (n = 36) belonged to the clonal complex (CC)2, one isolate belonged to CC1, and one isolate belonged to CC164. Sixteen new STs (ST1235-ST1250) were identified. Identifying both global ST2 and a large number of new STs, revealed high genetic diversity in A. baumannii isolates in the study population.

[The Identification of Acinetobacter baumannii Blood Isolates by MALDI-TOF MS, ARDRA and blaOXA-51-like Gene-Specific Real-Time PCR]. / Acinetobacter baumannii Kan Izolatlarinin MALDI-TOF MS, ARDRA ve blaOXA-51-benzeri Gene Özgül Gerçek Zamanli Polimeraz Zincir Reaksiyonu ile Tanimlanmasi.

The Acinetobacter calcoaceticus-Acinetobacter baumannii (Acb) complex consists of phenotypically very similar nosocomial species; A.baumannii, Acinetobacter nosocomialis, Acinetobacter pittii, Acinetobacter seifertii and Acinetobacter djikshoorniae and one environmental species A.calcoaceticus. The rapid and accurate identification of the members of Acb complex is critical as these nosocomial pathogens can show differences in antimicrobial susceptibility and clinical outcomes. The conventional phenotypic methods are slow, unreliable and less efficient for the differentiation of Acb complex species, including the A.baumannii species within the Acb complex. Although various molecular methods are available, such as amplified ribosomal DNA restriction analysis (ARDRA) and blaOXA-51-like gene specific PCR, they are usually inconvenient for the routine diagnostic laboratories. Recently, matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) offers an opportunity for rapid, cost-effective and convenient bacterial identification in routine diagnostic procedures conducted in clinical laboratory. In this study, we aimed to evaluate the diagnosis performance of MALDI-TOF MS system to identify blood isolates of A.baumannii. A total of 180 nonduplicate carbapenem resistant Acb complex (strain numbers; TR1-TR60) and A.baumannii (TR61-TR180) blood isolates were collected from the intensive care units of the three university hospitals in Turkey from January 2016 to December 2016. All isolates were evaluated by using blaOXA-51-like gene specific real time (Rt-PCR) analysis, ARDRA (restriction enzymes-AluI, CfoI, MboI, MspI, RsaI) method and MALDI-TOF MS (VITEK® MS, bioMérieux, France) system. All the strains except TR10, TR31, TR35 and TR52 were identified as A.baumannii by ARDRA method. Out of 177 of all the isolates, presence of blaOXA-51-like gene was found except for TR10, TR31 and TR52 isolates. However, TR31 without the presence of blaOXA-51-like gene was identified as A.pittii using the ARDRA. Totally 176 isolates which were identified as A.baumannii by both of the methods, ARDRA and Rt-PCR- blaOXA-51-like, were accepted as a reference for the evaluation of the diagnosis performance capacity of the MALDI-TOF MS. Overall, for all 176 isolates tested, the sensitivity obtained with the MALDI-TOF MS were 99.4% with 75% specificity. The accuracy value of the method was determined as 98.9% for the identification of A.baumannii to the species level. MALDI-TOF MS is increasingly used in diagnostic microbiology for the routine identification of bacteria to the genus, species or subspecies level with high rates of sensitivity and specificity. In future, by expanding the database, MALDI-TOF MS system would possibly become the ideal method for routine diagnostic laboratories that could potentially identify more species and even determine some characteristics of antimicrobial resistance and virulence determinants.

Efectos de la vacuna neumocócica conjugada (VCN7 y VCN13) en los niños turcos con enfermedad neumocócica invasiva: experiencia en un solo centro / The effects of pneumococcal conjugate vaccine (PCV7 and PCV13) on Turkish children with invasive pneumococcal disease: a single center experience

Introducción: Nuestro objetivo fue determinar los cambios en la incidencia de enfermedad neumocócica invasiva (ENI), la distribución de serotipos y patrones de resistencia antibiótica del Streptococcus pneumoniae en niños con ENI tras el período de vacunación (de 1 a 7 años) con vacuna neumocócica de 7 serotipos (VCN7) (2008) y de 13 serotipos (VCN13) (2011). Población y métodos: El estudio se realizó en 39 niños con ENI de 1 mes a 18 años de edad en Angora, Turquía. Se identificó Streptococcus pneumoniae en sangre, líquido cefalorraquídeo, líquido pleural, y otros tejidos y líquidos corporales estériles mediante procedimientos estándar. Se analizó la resistencia de cepas aisladas de S. pneumoniae a penicilina y ceftriaxona con la prueba de epsilometría (E-test). Los serotipos de las cepas se determinaron con la reacción de Quellung. Resultados: La incidencia anual de ENI disminuyó significativamente de 7,71 (intervalo de confianza --#91;IC--#93; del 95%: de 1,99 a 13,4) a 1,58 (IC del 95%: de 0,6 a 3,77; reducción del riesgo relativo= -79,5; p= 0,006) cada 100 000 habitantes de < 5 años de edad sin enfermedad preexistente. Durante todo el período del estudio, los serotipos en la VCN7 y en la VCN13 representaron el 27,8% y el 63,8% de las cepas aisladas, respectivamente. Los serotipos en la VCN13 correspondían al 81,8% de los casos de ENI en la era previa a la introducción de esta vacuna, y disminuyeron al 56% en los cuatro años posteriores. Las tasas de resistencia a penicilina y ceftriaxona (en el caso de la meningitis) fueron del 48,5% y el 9,1%, respectivamente. Conclusiones: Este estudio observó una disminución significativa en la incidencia de ENI después de la introducción de la VCN13.

Introduction. The aim of this prospective singlecenter study was to determine the changings in incidence of invasive pneumococcal disease (IPD), serotype distribution and the antimicrobial resistance patterns of S. pneumoniae in children with IPD after the period (1 to 7 years) of vaccination with PCV7 (2008) and PCV13 (2011). Population and methods. The study was conducted on 39 Turkish children with IPD between ages 1 month and 18 years in Ankara, Turkey. Streptococcus pneumoniae was identified using standard laboratory procedures from blood, cerebrospinal fluid (CSF), pleural fluid, and other sterile body fluids and tissues. S. pneumoniae isolates were tested for resistance to penicilin and ceftriaxone using the E-test methodology. Serotypes of the isolates were determined by Quellung reaction. Results. The overall annual incidence rate of IPD decreased significantly from 7.71 (95% CI, 1.99-13.4) to 1.58 (95% CI, 0.6-3.77; RRR= -79.5; p= 0.006) per 100 000 population among <5 years of age without underlying disease. During the overall study period, the PCV7-serotypes and PCV13-serotypes represented 27.8% and 63.8% of isolates, respectively. PCV13-serotypes made up 81.8% of cases of IPD in the pre-PCV13 era and decreased to 56% in the 4 years after PCV13. The penicillin and ceftriaxone (for meningitis) resistance rates were 48.5% and 9.1%, respectively. Conclusions. This is the first study about the changing pattern of the incidence of IPD in Turkish children after the implementation of the PCV7 and PCV13 in Turkish national vaccine schedule and a prominent decrease in incidence of IPD has seen after the implementation of PCV13.

The effects of pneumococcal conjugate vaccine (PCV7 and PCV13) on Turkish children with invasive pneumococcal disease: a single center experience. / Efectos de la vacuna neumocócica conjugada (VCN7 y VCN13) en los niños turcos con enfermedad neumocócica invasiva: experiencia en un solo centro.

INTRODUCTION: The aim of this prospective single-center study was to determine the changings in incidence of invasive pneumococcal disease (IPD), serotype distribution and the antimicrobial resistance patterns of S. pneumoniae in children with IPD after the period (1 to 7 years) of vaccination with PCV7 (2008) and PCV13 (2011). POPULATION AND METHODS: The study was conducted on 39 Turkish children with IPD between ages 1 month and 18 years in Ankara, Turkey. Streptococcus pneumoniae was identified using standard laboratory procedures from blood, cerebrospinal fluid (CSF), pleural fluid, and other sterile body fluids and tissues. S. pneumoniae isolates were tested for resistance to penicilin and ceftriaxone using the E-test methodology. Serotypes of the isolates were determined by Quellung reaction. RESULTS: The overall annual incidence rate of IPD decreased significantly from 7.71 (95% CI, 1.99-13.4) to 1.58 (95% CI, 0.6-3.77; RRR=-79.5; p=0.006) per 100 000 population among ≤5 years of age without underlying disease. During the overall study period, the PCV7-serotypes and PCV13-serotypes represented 27.8% and 63.8% of isolates, respectively. PCV13-serotypes made up 81.8% of cases of IPD in the pre-PCV13 era and decreased to 56% in the 4 years after PCV13. The penicillin and ceftriaxone (for meningitis) resistance rates were 48.5% and 9.1%, respectively. CONCLUSIONS: This is the first study about the changing pattern of the incidence of IPD in Turkish children after the implementation of the PCV7 and PCV13 in Turkish national vaccine schedule and a prominent decrease in incidence of IPD has seen after the implementation of PCV13.

INTRODUCCIÓN: Nuestro objetivo fue determinar los cambios en la incidencia de enfermedad neumocócica invasiva (ENI), la distribución de serotipos y patrones de resistencia antibiótica del Streptococcus pneumoniae en niños con ENI tras el período de vacunación (de1 a 7 años) con vacuna neumocócica de 7 serotipos (VCN7) (2008) y de 13 serotipos (VCN13) (2011). POBLACIÓN Y MÉTODOS: El estudio se realizó en 39 niños con ENI de 1 mes a 18 años de edad en Angora, Turquía. Se identificó Streptococcus pneumoniae en sangre, líquido cefalorraquídeo, líquido pleural, y otros tejidos y líquidos corporales estériles mediante procedimientos estándar.Se analizó la resistencia de cepas aisladas de S. pneumoniae a penicilina y ceftriaxona con la prueba de epsilometría (E-test). Los serotipos de las cepas se determinaron con la reacción de Quellung. RESULTADOS: La incidencia anual de ENI disminuyó significativamente de 7,71 (intervalo de confianza [IC] del 95%: de 1,99 a 13,4) a 1,58 (IC del 95%: de 0,6 a 3,77; reducción del riesgo relativo=-79,5; p=0,006) cada 100 000 habitantes de ≤ 5 años de edad sin enfermedad preexistente. Durante todo el período del estudio, los serotipos en la VCN7 y en la VCN13 representaron el 27,8% y el 63,8% de las cepas aisladas, respectivamente. Los serotipos en la VCN13 correspondían al 81,8% de los casos de ENI en la era previa a la introducción de esta vacuna, y disminuyeron al 56% en los cuatro años posteriores. Las tasas de resistencia a penicilina y ceftriaxona (en el caso de la meningitis) fueron del 48,5% y el 9,1%, respectivamente. CONCLUSIONES: Este estudio observó una disminución significativa en la incidencia de ENI después de la introducción de la VCN13.

Comparison of in-house biotin-avidin tetanus IgG enzyme-linked-immunosorbent assay (ELISA) with gold standard in vivo mouse neutralization test for the detection of low level antibodies.

BACKGROUND: Detection of anti-tetanus antibody levels is necessary for both determination of the immune status of individuals and also for planning preventive measures. ELISA is the preferred test among in vitro tests however it can be affected by the cross reacting antibodies. A previously developed in-house ELISA test was found not reliable for the antibody levels ≤1.0IU/ml. A new method was developed to detect low antibody levels correctly. The aim of the present study was to compare the results of the newly developed in-house biotin-avidin tetanus IgG ELISA test with the in vivo mouse neutralization test, for the antibody levels ≤1.0IU/ml. METHODS: A total of 54 serum samples with the antibody levels of three different levels, =0.01IU/ml, 0.01-0.1IU/ml, 0.1-1IU/ml, which were detected by in vivo mouse neutralization test were studied by the newly developed in-house biotin-avidin tetanus IgG ELISA test. Test was validated by using five different concentrations (0.01IU/ml, 0.06IU/ml, 0.2IU/ml, 0.5IU/ml, 1.0IU/ml). RESULTS: A statistically significant correlation (r2=0.9967 p=0,001) between in vivo mouse neutralization test and in-house biotin-avidin tetanus IgG ELISA test, was observed. For the tested concentrations intra-assay, inter-assay, accuracy, sensitivity, specificity and coefficients of variations were determined as ≤15%. CONCLUSION: In-house biotin-avidin tetanus IgG ELISA test can be an alternative method to in vivo mouse neutralization method for the detection of levels ≤1.0IU/ml. By using in-house biotin-avidin tetanus IgG ELISA test, individuals with non protective levels, will be reliably detected.

Ureaplasma urealyticum: Presence among Sexually Transmitted Diseases.

The aim of this study was to detect the presence of Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium, Mycoplasma hominis, Trichomonas vaginalis, and Ureaplasma urealyticum in genital specimens of symptomatic patients. This study also examined the role of U. urealyticum in infections of the lower genital tract. Cervical and urethral samples from 96 patients (46 males, 50 females) were tested using the Seeplex(®) STD6 ACE kit. Consent forms were received and a questionnaire was applied. All statistical analyses were performed using the SPSS statistical software program (version 17.0). Among the samples tested, at least 1 pathogen was detected in 49% of the samples; specifically, the rate of detection of U. urealyticum, M. hominis, C. trachomatis, N. gonorrhoeae, M. genitalium, and T. vaginalis was 29.1%, 10.4%, 8.3%, 7.3%, 6.3%, and 4.2%, respectively. U. urealyticum was detected as the sole pathogen in samples from 10% of female patients and 28.3% of male patients (p = 0.035). U. urealyticum was present in 54.5% (18/33) of samples in which a single pathogen was detected and 71.4% (10/14) of samples in which multiple pathogens were detected. Among men, significant differences in discharge, dysuria, and pruritus were not noted among those with negative results (84.6%, 69.2%, and 38.5%, respectively), among those positive for only U. urealyticum (100%, 66.7%, and 26.7%, respectively), and those positive for N. gonorrhoeae, C. trachomatis, M. genitalium, and T. vaginalis (100%, 93.3%, and 26.7%, respectively). Detection of U. urealyticum, either alone or together with other pathogens, in a symptomatic group of patients is an important finding, particularly in men.

Comparison of a novel real-time PCR method (RTA) and Artus RG for the quantification of HBV DNA and HCV RNA.

INTRODUCTION: The purpose of this study was to evaluate and compare two manual isolation and real-time (RT) polymerase chain reaction (PCR) kits (RTA RT-PCR with RTA isolation kit and Artus RG RT-PCR with QIAamp isolation kit) for molecular diagnosis of hepatitis B virus (HBV) and hepatitis C virus (HCV) infections. METHODOLOGY: The study was conducted on 121 and 54 clinical samples for the detection of HBV DNA and HCV RNA, respectively, with an additional 8 HCV RNA external quality control samples. RESULTS: Though a high correlation was observed between the two kits for the HBV DNA (r = 0.955, p = 0.001) and HCV RNA quantifications (r = 0.828, p = 0.001), discordant results were found in nine of the HBV DNA and in six of the HCV RNA samples. The mean difference between the two systems was found to be 0.4 log IU/mL in Quality Control for Molecular Diagnostics (QCMD) HCV RNA samples by Bland-Altman analysis. CONCLUSIONS: Although there was a high correlation between HBV DNA and HCV RNA tests according to the results of the study, the RTA system requires improvement for the determination of HCV RNA.

[Serological evaluation of Bordetella pertussis infection in adults with prolonged cough]. / Uzamis öksürügü olan eriskinlerde Bordetella pertussis enfeksiyonunun serolojik olarak degerlendirilmesi.

Pertussis is a vaccine-preventable disease that is transmitted from infected to susceptible individuals by respiratory route. Bordetella pertussis infection may occur at any age as neither vaccine nor natural infection induced immunity lasts life-long. This study was planned to demonstrate the serological evidence of infection among adults, to raise awareness among clinicians and to provide data for the development of strategies to protect vulnerable infants. A total of 538 patients (345 female, 193 male) ages between 18-87 years who had a complain of prolonged cough for more than two weeks were included in the study. Anti-pertussis toxin (PT) IgG and anti-filamentous hemagglutinin (FH) IgG levels from single serum samples were measured by an in-house ELISA test which was standardized and shown to be efficient previously. Anti-PT IgG antibody levels of ≥ 100 EU/ml were considered as acute/recent infection with B.pertussis. In our study, 9.7% (52/538) of the patients had high levels of anti-PT IgG (≥ 100 EU/ml) and among those patients 43 (43/52; 82.7%) also had high (≥ 100 EU/ml) anti-FHA IgG levels. There were no statistically significant differences in terms of age, gender, education level, DPT (diphtheria-pertussis-tetanus) vaccination history, smoking history or average daily cigarette consumption (p> 0.05) between the cases with high antibody levels (n= 52). When the symptoms and the presence of cases with high antibody levels were evaluated, it was detected that no one parameter was significantly different from others, except that 24.1% of the cases with inspiratory whooping had high anti-PT levels. There was also no statistically significant difference between high anti-PT levels ≥ 100 EU/ml and the patients with risk factors [smoking (21/200; 10.5%), presence of disease that cause chronic cough and/or drug usage (19/171; %11.1), and whole factors which cause chronic cough (32/306; %10.5)] and without risk factors (p= 0.581; p= 0.357; p= 0.249, respectively). The distribution of anti-PT IgG geometric mean titer (GMT) according to the age groups, was as follows; 32.41 in 18-30 years; 36.28 in 31-50 years; 36.82 in 51-70 years and 31.15 in ≥ 71 years. Our results indicated that B.pertussis infections are also present among adult population with a frequency not to be underestimated (9.7%) and the results also emphasized that since typical whooping cough symptoms may not be seen in adults, pertussis infection should be considered as a differential diagnosis in adults with prolonged cough, even if there are some other underlying factors of cough. The data obtained from this study was also considered to be helpful in the development of adult vaccination policies for the protection of infants who have not completed the vaccination schedule yet.

The correlation of sodium iodide symporter and BRAF(V600E) mutation in classical variant papillary thyroid carcinoma.

BRAF(V600E) mutation was analyzed by real-time polymerase chain reaction in 96 consecutive cases with classical variant papillary thyroid cancer, and immunohistochemical staining of Na+/I- symporter (NIS) protein was evaluated. Localization (intracellular or membranous), density, and the intensity of cytoplasmic staining were characterized semiquantitatively. Extrathyroidal invasion, surgical margin positivity, and lymph node metastasis were compared with BRAF(V600E) mutation and NIS expression. Eighty-eight patients who had at least 24-month follow-up were also included in survival analysis. BRAF(V600E) mutation was determined in 78.1% (75/96) and functional NIS activity in 74% (71/96) of the cases. There were statistically significant differences in mean ages between BRAF(V600E) mutation-positive (48.6) and BRAF(V600E) mutation-negative cases (37.3; Levene test, P=.419; Student t test, P=.001). The surgical margin positivity (46.7%) and extrathyroidal extension percentage (54.7%) in the BRAF(V600E) mutation-positive group were higher than the negative (28.6% and 33.3%, respectively) group, without statistical significance (P=.138 and P=.084, respectively). Functional NIS activity was higher in BRAF(V600E) mutation-positive cases (78.1%) than mutation-negative ones (57.1%; P=.047). The possibility of moderate and intense cytoplasmic staining in BRAF(V600E) mutation-positive cases (72%) was 6.3 times higher than the possibility of weak staining (28%) in the mutation-positive cases (95% confidence interval, 2.2-18.8; P=.001). Functional NIS expression is higher in patients with classical variant papillary thyroid cancer with BRAF(V600E) mutation. However, the clinical features were not found to be associated with NIS expression. There may be different mechanisms determining the outcome of therapy.

High-Level Genetic Diversity among Invasive Streptococcus pneumoniae Isolates in Turkey.

This study obtained information on the serotypes and molecular typing characteristics of Streptococcus pneumoniae strains causing invasive diseases in Turkey. Sixty-eight S. pneumoniae isolates causing invasive pneumococcal diseases were collected from different regions of Turkey from 2009 to 2011. The isolates were characterized by performing multilocus sequence typing (MLST), pulsed-field gel electrophoresis (PFGE), and capsular serotyping, and 25 different serotypes were identified. Serotypes 19F, 23F, 1, 14, and 7F were common and accounted for 52.9% of all the serotypes. In addition, 54 different PFGE profiles (pulsotypes) were observed. Twenty-three of the 68 (33.8%) isolates were clustered into 9 pulsotypes. MLST analysis yielded 36 sequence types, of which 12 (33.3%) were novel. A comparison of results with the global pneumococcal MLST database by performing eBURST analysis showed that our strains belonged to 20 different clonal complexes and 5 singletons. In addition, we identified 4 new alleles: 2 gdh, 1 xpt, and 1 ddl. Thus, the results of this study highlighted a high level of diversity among pneumococcal isolates. In addition, the study identified a case of possible capsular switching.

[Extended-spectrum beta-lactamase production by Enterobacteriaceae isolates from urine cultures of outpatients: results of a 7-year follow-up]. / Ayaktan basvuran hastalarin idrar kültürlerinde üretilen Enterobacteriaceae izolatlarinda genislemis spektrumlu beta-laktamaz üretimi: 7 yillik izlem sonuçlari.

The aim of the study was to evaluate the change of the frequency of extended-spectrum beta-lactamase (ESBL) producing Enterobacteriaceae isolates from urine samples of outpatients in years and to analyse the antibiotic resistance profiles for a rational drug use. The urine samples cultured in our laboratory from the patients who were admitted to outpatient clinics of our hospital between years 2007-2013 were included in this study. Enterobacteriaceae strains were isolated and identified by conventional methods and API 20E system (BioMérieux, France). The standard antimicrobial susceptibility tests were performed by Kirby Bauer disk diffusion method. ESBL production were screened by double-disk synergy method according to CLSI guidelines. E-test method (BioMérieux, France) were used for the verification of suspicious ESBL production. The identification and antimicrobial susceptibility testing were performed for a total of 12.535 isolates. Of the isolates 8716 were identified as Escherichia coli (69.3%), 1514 were Klebsiella pneumoniae/oxytoca (12.1%), 257 were Proteus mirabilis (2.1%), 345 were other Enterobacteriae members (8%), 411 were various non-fermentative gram-negative bacteria (3.3%) and 1292 were various gram-positive bacteria (10.3%). The total positivity rate of ESBL was found as 21.8% (2.283/10.487), and the ESBL positive rates for E.coli, K.pneumoniae/oxytoca and P.mirabilis were 21.2%, 28.2% and 4.7%, respectively. Other Enterobacteriaceae isolates were not evaluated because of the absence of standardized methods and breakpoint values. There was no statistically significant difference among ESBL producing isolates within seven years (p= 0.364). The antibiotic resistance rates of the ESBL-positive isolates were statistically higher than ESBL-negative isolates [amoxicillin-clavulanate (73.1%/11.3%), trimethoprim-sulfamethoxazole (63.1%/31.0%), nitrofurantoin (17.3%/8.6%), gentamicin (42.2%/10.1%), amikacin (3.5%/0.9%), tobramisin (56.8%/10.5%), imipenem (0.3%/0.1%), ofloxacin (66.8%/19.8%), ticarcillin-clavulanate (73.5%/19.8%), piperacillin-tazobactam (28.8%/5.0%)] (p< 0.05). Statistically significant variations were detected within the years for the resistance rates of amoxicillin-clavulanate (p= 0.001), tobramycin (p=0.003), ofloxacin (p= 0.001), ticarcillin-clavulanate (p= 0.001) and piperacillin-tazobactam (p= 0.001) were detected within the years. Although a quite high percentage of ESBL positivity in Enterobacteriaceae isolates was determined, there was a slight but not statistically significant increase of this value during the seven-year period. The stability of the percentage of ESBL positivity may indicate a positive change in the habit of the usage of beta-lactam antibiotics. According to the results of our study, the most effective drugs for ESBL-producing isolates were piperacillin-tazobactam among inhibitor combinations, amikacin among aminoglycosides and nitrofurantoin among orally-used drugs.

Molecular characterization of vancomycin-resistant Enterococcus faecium strains isolated from carriage and clinical samples in a tertiary hospital, Turkey.

This study aimed to determine the presence of vancomycin resistance (vanA and vanB) and virulence genes (esp, asa1, gelE, ace, hyl, cylA, cpd and ebpA) in vancomycin-resistant Enterococcus faecium (VREfm) strains and to analyse the clonal relationships among the strains. E. faecium strains were identified from rectal and clinical specimens by biochemical tests and the API-20 Strep kit. Susceptibility testing was performed using disc-diffusion and broth-dilution methods. PFGE was used for molecular typing of the VREfm strains. The vancomycin resistance and virulence genes were amplified by two-step multiplex PCR. All 55 VREfm isolates were resistant to penicillin G, ampicillin and high-level gentamicin but were susceptible to quinupristin/dalfopristin and linezolid. Multiplex PCR analysis indicated that all isolates harboured vanA and that 41 (75 %) were positive for virulence genes. The esp gene was the most common virulence factor and was detected in nine (41 %) invasive and 32 (96.7 %) non-invasive isolates. Multiple virulence genes were observed only in two non-invasive isolates; one harboured esp and ebpA and the other harboured esp, ebpA, asa1, gelE and cpd. PFGE typing yielded 16 different types, seven of which were clusters with two to 14 strains each. The clustering rates of the rectal swab, blood and urine isolates were 72.7 %, 61.5 % and 87.5 %, respectively. The genetic similarity observed among the VREfm isolates indicated cross-transmission in the hospital. Further studies on the virulence factors present in the strains might provide insight into the acquisition of these traits and their contribution to increased prevalence of VREfm.

Serologic Investigation of Hantavirus Infection in Patients with Previous Thrombocytopenia, and Elevated Urea and Creatinine Levels in an Epidemic Region of Turkey.

The first cases of Hantavirus infection in Turkey were reported in early 2009 in the Zonguldak and Bartin provinces. The aim of this study was to investigate the presence of Hantavirus antibodies in patients who had clinical and laboratory findings that were potentially associated with Hantavirus infection prior to the epidemic in Bartin in 2009. After screening 314,577 medical records from between 2007 and 2009, the clinical and laboratory data for 442 patients meeting the criteria of coexistent thrombocytopenia, and elevated urea and creatinine levels were transferred to a statistical program. Home visits were made to 170 patients, 84 of whom consented to participate in the study. The participants completed a questionnaire and provided a blood sample. Commercial anti-Hantavirus IgG and IgM ELISA and immunoblotting assays were used, with seropositive samples being confirmed by focus reduction neutralization tests (FRNT). ELISA and/or immunoblotting assays detected 10 positive samples; however, only 7 of these were recorded as positive by FRNT. FRNT positivity was significantly associated with female sex, the presence of a barn near to the house, and working in a forest (P < 0.05). In a Hantavirus endemic region, physicians must keep in mind that thrombocytopenia, and elevated urea and creatinine levels may indicate Hantavirus infection.