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OBJECTIVES: To evaluate the restart of the German Bundesliga (football (soccer)) during the COVID-19 pandemic from a medical perspective. METHODS: Participants were male professional football players from the two highest German leagues and the officials working closely with them. Our report covers nine match days spread over 9 weeks (May to July 2020). Daily symptom monitoring, PCR testing for SARS-CoV-2 RNA twice weekly, and antibody tests (on two occasions-early during the phase in May 2020 and in the week of the last match) were conducted. Target variables were: (1) onset of typical COVID-19 symptoms, (2) positive PCR results, and (3) IgG seroconversion against SARS-CoV-2. All detected seroconversions were controlled by neutralisation tests. FINDINGS: Suspicious symptoms were reported for one player; an immediate additional PCR test as well as all subsequent diagnostic and antibody tests proved negative for coronavirus. Of 1702 regularly tested individuals (1079 players, 623 officials members), 8 players and 4 officials tested positive during one of the first rounds of PCR testing prior to the onset of team training, 2 players during the third round. No further positive results occurred during the remainder of the season. 694 players and 291 officials provided two serum samples for antibody testing. Nine players converted from negative/borderline to positive (without symptoms); two players who initially tested positive tested negative at the end of the season. 22 players remained seropositive throughout the season. None of the seroconversions was confirmed in the neutralisation test. CONCLUSION: Professional football training and matches can be carried out safely during the COVID-19 pandemic. This requires strict hygiene measures including regular PCR testing.
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Prueba de Ácido Nucleico para COVID-19 , COVID-19/diagnóstico , Volver al Deporte , SARS-CoV-2 , Fútbol/estadística & datos numéricos , Adulto , Anticuerpos Antivirales/sangre , COVID-19/epidemiología , COVID-19/prevención & control , Prueba de Ácido Nucleico para COVID-19/estadística & datos numéricos , Estudios de Cohortes , Alemania/epidemiología , Humanos , Inmunoglobulina G/sangre , Masculino , Pruebas de Neutralización , Estudios Prospectivos , SARS-CoV-2/inmunología , Seguridad , Evaluación de Síntomas/métodosRESUMEN
The Lower Austrian Wachau region was an early COVID-19 hotspot of infection. As previously reported, in June 2020, after the first peak of infections, we determined that 8.5% and 9.0% of the participants in Weißenkirchen and surrounding communities in the Wachau region were positive for immunoglobulin G (IgG) and immunoglobulin A (IgA) antibodies against the receptor-binding domain of the spike protein of SARS-CoV-2, respectively. Here, we present novel data obtained eight months later (February 2021) from Weißenkirchen, after the second peak of infection, with 25.0% (138/552) and 23.6% (130/552) of participants that are positive for IgG and IgA, respectively. In participants with previous IgG/IgA positivity (June 2020), we observed a 24% reduction in IgG levels, whereas the IgA levels remained stable in February 2021. This subgroup was further analyzed for SARS-CoV-2 induced T cell activities. Although 76% (34/45) and 76% (34/45) of IgG positive and IgA positive participants, respectively, showed specific T cell activities (upon exposure to SARS-CoV-2 spike protein-derived peptides), those were not significantly correlated with the levels of IgG or IgA. Thus, the analyses of antibodies cannot surrogate the measurement of T cell activities. For a comprehensive view on SARS-CoV-2-triggered immune responses, the measurement of different classes of antibodies should be complemented with the determination of T cell activities.
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Formación de Anticuerpos , COVID-19 , Glicoproteína de la Espiga del Coronavirus , Anticuerpos Antivirales , Austria/epidemiología , COVID-19/epidemiología , COVID-19/inmunología , Humanos , Inmunoglobulina A , Inmunoglobulina G , Inmunoglobulina M , SARS-CoV-2 , Glicoproteína de la Espiga del Coronavirus/inmunologíaRESUMEN
PURPOSE: We aimed to elaborate whether cycle threshold (Ct) values differ significantly between wild type SARS-CoV-2 (wtV) and certain viral variants and how strong or weak a potential significant effect might be. METHODS: In a retrospective study, we investigated 1873 SARS-CoV-2 positive samples for the occurrence of viral marker mutations. Age, gender, clinical setting, days after onset of symptoms, and Ct values were recorded. Statistical analysis was carried out with special consideration of effect sizes. RESULTS: During the study period wtV was detected in 1013 samples (54%), while 845 (45%) patients carried the Alpha variant of concern (VOC), and 15 (1%) the Beta VOC. For further analysis, only wtV and the Alpha VOC were included. In a multi-factor ANOVA and post-hoc test with Bonferroni-correction for the age groups we found significant main-effects for Ct values of the viral variant (wtV mean 26.4 (SD 4.27); Alpha VOC mean 25.0 (SD 3.84); F (1,1850) = 55.841; p < .001) and the clinical setting (outpatients: mean 25.7 (SD 4.1); inpatients: mean 27.0 (SD 4.2); F (1,1850) = 8.520, p = .004). However, since the effect sizes were very small (eta squared for the Alpha VOC = .029 and the clinical setting = .004), there was only a slight trend towards higher viral loads of the Alpha VOC compared to wtV. CONCLUSIONS: In order to compare different variants of SARS-CoV-2 the calculation of effect sizes seems to be necessary. A combination of p-values as estimates of the existance of an effect and effect sizes as estimates of the magnitude of a potential effect may allow a better insight into transmission mechanisms of SARS-CoV-2.
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COVID-19 , SARS-CoV-2 , Humanos , Estudios Retrospectivos , SARS-CoV-2/genética , Pruebas SerológicasRESUMEN
Background: Since December 2019 the novel coronavirus (SARS-CoV-2) is the center of global attention due to its rapid transmission and toll on health care systems and global economy. Population-based serosurveys measuring antibodies for SARS-CoV-2 provide one method for estimating previous infection rates including the symptom-free courses of the disease and monitoring the progression of the epidemic. Methods: In June 2020 we succeeded in testing almost half of the population of an Austrian township (1,359 inhabitants) with a reported higher incidence for COVID-19 infections (17 PCR positive cases have been officially reported until the date of sample collection, i.e., 1.2% of the total population). We determined the prevalence of SARS-CoV-2-specific antibodies in this population, factors affecting, and symptoms correlated with prior infection. Antibodies were determined using a CE-certified quality-controlled ELISA test for SARS-CoV-2-specific IgG and IgA antibodies. Results: We found a high prevalence of 9% positive antibodies among the town population in comparison to 6% of the neighboring villages. This was considerably higher than the officially known RT-PCR-approved COVID-19 cases (1.2%) in the town population. Twenty percent of SARS-CoV-2-antibody positive cases declared being asymptomatic in a questionnaire. On the other hand, we identified six single major symptoms, including anosmia/ageusia, weight loss, anorexia, general debility, dyspnea, and fever, and especially their combination to be of high prognostic value for predicting SARS-CoV-2 infection in a patient. Conclusions: This population study demonstrated a high prevalence of antibodies to SARS-CoV-2 as a marker of past infections in an Austrian township. Several symptoms revealed a diagnostic value especially in combination.
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Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) caused a global pandemic recently. The prevalence and persistence of antibodies following a peak SARS-CoV-2 infection provides insights into the potential for some level of population immunity. In June 2020, we succeeded in testing almost half of the population of an Austrian town with a higher incidence of COVID-19 infection. We performed a follow-up study to reassess the prevalence of SARS-CoV-2-specific IgA and IgG antibodies with 68 participants of the previous study. We found that the prevalence of IgG or IgA antibodies remained remarkably stable, with 84% of our cohort prevailing SARS-CoV-2-specific antibodies (only a slight decrease from 93% 4 months before). In most patients with confirmed COVID-19 seroconversion potentially provides immunity to reinfection. Our results suggest a stable antibody response observed for at least 6 months post-infection with implications for developing strategies for testing and protecting the population.
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BACKGROUND: Measures of distancing, wearing face/medical masks and lockdown introduced in many countries to meet the challenges of the SARS-CoV-2 pandemic have led to gross changes in the epidemiology of important infections. The observation of decline of positive norovirus tests after introduction of lockdown in Germany led us to investigate changes in the detection of major causes of diarrhoea by comparing pre-pandemic quarters (PPQ: 1Q/17 through 1Q/20) since 2017 and pandemic quarters (PQ: 2Q/20 through 1Q/21). METHODS AND SETTING: Bioscientia Laboratory Ingelheim is a large regional clinical pathology laboratory serving > 50 hospitals and > 5000 general practitioners and specialist outpatient practices located in the federal states Hesse, Rhineland-Palatinate and North Rhine-Westphalia, Germany. Antigen detection assays were used for detection of astrovirus, adenovirus, rotavirus, and Campylobacter antigen and Clostridium difficile Toxin A/B, while norovirus was detected by qualitative RT-PCR. FINDINGS: The mean positivity-ratios of norovirus, adenovirus and astrovirus assays were 3-20 fold lower in periods PQ (2Q/20 through 1Q/21) compared to PPQ (1Q/17 through 1Q/20) (p<.01). The mean positivity-ratio was lower in PQ compared to PPQ for rotavirus (p=.31), but failed to reach statistical significance, while for campylobacter antigen (p=.91) and C. difficile Toxin A/B (p=.17) the mean positivity-ratio was even higher in PQ compared to PPQ. CONCLUSIONS: Apparently, hygienic measures used to contain the SARS-CoV-2 pandemic have differential effects on incidence of diarrhoea viruses as compared to bacterial gastrointestinal agents, particularly C. difficile, which may lead to re-evaluate measures implemented against this important cause of nosocomial diarrhoea.
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OBJECTIVES: Elite professional football players and staff are a unique group that might give insight into the epidemiology of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections in Germany and thus can serve as a model for geographical distribution and an estimation of undetected infections. METHODS: In this prospective cohort study seroprevalence was determined twice in May and June 2020 in players and staff from the German Bundesliga. As screening assays, a commercial ELISA (Euroimmun) and a chemiluminescent immunoassay (CLIA) (Roche) were used, and an in-house neutralization assay (NT) was used as reference standard. Participants were tested twice weekly using PCR from nasopharyngeal and/or oropharyngeal swabs. RESULTS: Seroprevalence (NT used as confirmation) in 2164 samples from 1184 players and staff was rather similar in May (23/1157, 1.99%) and June (21/1007, 2.09%). All participants were PCR-negative during the study period. Significant regional differences in seroprevalence were not observed. When comparing seroprevalence with the cumulative incidence of infections derived from the German notification system (subgroup matching to cohort; men, age 20-69 years), IgG was found eight to ten times more frequently, pointing to a high rate of undetected infection. ELISA and CLIA correlated only moderately (κ 0.52). CONCLUSIONS: Seroprevalence with a high-quality diagnostic in Germany seemed to be around 2%. The number of undetected infections seems to be eight to ten times higher than in notification data. The quality of antibody assays is rather variable, thus results should ideally be confirmed at least by a second assay to prove IgG positivity.
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Infecciones Asintomáticas , COVID-19 , Inmunoglobulina G , SARS-CoV-2 , Fútbol , Adulto , Anciano , Humanos , Masculino , Persona de Mediana Edad , Adulto Joven , Anticuerpos Antivirales/sangre , Infecciones Asintomáticas/epidemiología , COVID-19/diagnóstico , COVID-19/epidemiología , Alemania/epidemiología , Inmunoensayo/métodos , Inmunoglobulina G/sangre , Incidencia , Reacción en Cadena de la Polimerasa , Prevalencia , Estudios Prospectivos , ARN Viral/genética , SARS-CoV-2/inmunología , SARS-CoV-2/aislamiento & purificación , Estudios SeroepidemiológicosRESUMEN
BACKGROUND: Amplification of viral ribonucleic acid (RNA) by real-time reverse transcriptase polymerase chain reaction (rRT-PCR) is the gold standard to detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Since the initial outbreak, strategies to detect and isolate patients have been important to avoid uncontrolled viral spread. Although testing capacities have been upscaled, there is still a need for reliable high throughput test systems, specifically those that require alternative consumables. Therefore, we tested and compared two different methods for the detection of viral PCR products: rRT-PCR and mass spectrometry (MS). METHODS: Viral RNA was isolated and amplified from oro- or nasopharyngeal swabs. A total of 22 samples that tested positive and 22 samples that tested negative for SARS-CoV-2 by rRT-PCR were analyzed by MS. Results of the rRT-PCR and the MS protocol were compared. RESULTS: Results of rRT-PCR and the MS test system were in concordance in all samples. Time-to-results was faster for rRT-PCR. Hands-on-time was comparable in both assays. CONCLUSIONS: MS is a fast, reliable and cost-effective alternative for the detection of SARS-CoV-2 from oral and nasopharyngeal swabs.
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Betacoronavirus/aislamiento & purificación , Infecciones por Coronavirus/diagnóstico , Espectrometría de Masas/métodos , Neumonía Viral/diagnóstico , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , COVID-19 , Niño , Infecciones por Coronavirus/virología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Pandemias , Neumonía Viral/virología , ARN Viral/análisis , Reacción en Cadena en Tiempo Real de la Polimerasa , SARS-CoV-2 , Factores de Tiempo , Adulto JovenRESUMEN
AIMS: Strong evidence shows that late diabetic complications in diabetes mellitus are substantially related to an increased synthesis of diacylglycerol with a subsequent activation of protein kinase C (PKC) beta. Several studies have shown that specific inhibition of the PKC isoform beta by ruboxistaurin is able to attenuate the development of microvascular complications under diabetic conditions. The aim of this in vitro study was to investigate the effect of ruboxistaurin on glucose-induced adhesion of monocytes to endothelial cells, representing one of the first pivotal steps in the course of atherogenesis. METHODS: Human umbilical venous endothelial cells were isolated and cultured to confluence in microtiter plates. After coincubation with monocytes in the presence of 0, 10, or 400 ng ruboxistaurin to achieve PKC beta-specific and -unspecific PKC inhibition, cells were fixed and monocyte adhesion was determined by means of a standardized chemiluminescence assay. Expression of adhesion molecules (intercellular adhesion molecule-1, vascular cell adhesion molecule-1, and E-selectin) was also measured by chemiluminescence methods. RESULTS: Adhesion of monocytes to endothelial cells cultured under hyperglycemic conditions (27.7 mM glucose) was increased by 30.9 +/- 5.1% (p < 0.001) versus endothelial cells cultured under normoglycemic (NG) conditions (5.5 mM). Pretreatment of endothelial cells with 10 nM (PKC beta-specific concentration) and 400 nM (PKC beta-unspecific concentration) led to a significant reduction of glucose-induced adhesion of monocytes to endothelial cells that was statistically not different from endothelial adhesion under NG conditions (-7.2 +/- 3.1 and -8.1 +/- 2.6%, respectively; not significant vs NG). A nonsignificant tendency to lower the expression of adhesion molecules was seen with 10 ng of ruboxistaurin. CONCLUSIONS: We conclude that monocyte adhesion to endothelial cells under hyperglycemic conditions is at least mediated by PKC beta activation. Ruboxistaurin is able to suppress this monocyte adhesion even in a PKC beta-specific concentration. Further studies should evaluate these potential effects of ruboxistaurin in vivo.