There is an ongoing need for high-precision serological assays for the quantitation of anti-SARS-CoV-2 antibodies. Here, a trimeric SARS-CoV-2 spike (S) protein was used to develop an ELISA to quantify specific IgG antibodies present in serum, plasma, and dried blood spots (DBS) collected from infected patients or vaccine recipients. The quantitative S-ELISA was calibrated with international anti-SARS-CoV-2 immunoglobulin standards to provide test results in binding antibody units per mL (BAU/mL). The assay showed excellent linearity, precision, and accuracy. A sensitivity of 100% was shown for samples collected from 54 patients with confirmed SARS-CoV-2 infection more than 14 days after symptom onset or disease confirmation by RT-PCR and 58 vaccine recipients more than 14 days after vaccination. The assay specificity was 98.3%. Furthermore, antibody responses were measured in follow-up samples from vaccine recipients and infected patients. Most mRNA vaccine recipients had a similar response, with antibody generation starting 2–3 weeks after the first vaccination and maintaining positive for at least six months after a second vaccination. For most infected patients, the antibody titers increased during the second week after PCR confirmation. This S-ELISA can be used to quantify the seroprevalence of SARS-CoV-2 in the population exposed to the virus or vaccinated.

No description available.

The COVID-19 pandemic affected regular health care for patients with chronic diseases. However, the impact of the pandemic on primary care for patients with coronary artery disease (CAD) who are enrolled in a structured disease management program (DMP) in Germany is not clear. We investigated whether the pandemic affected primary care and health outcomes of DMP-CAD patients (n = 750) by using a questionnaire assessing patients’ utilization of medical care, CAD symptoms, as well as health behavior and mental health since March 2020. We found that out of concern about getting infected with COVID-19, 9.1% of the patients did not consult a medical practitioner despite having CAD symptoms. Perceived own influence on infection risk was lower and anxiety was higher in these patients compared to symptomatic CAD patients who consulted a physician. Among the patients who reported chest pain lasting longer than 30 min, one third did not consult a medical practitioner subsequently. These patients were generally more worried about COVID-19. Patients with at least one worsening CAD symptom (chest pain, dyspnea, perspiration, or nausea without apparent reason) since the pandemic showed more depressive symptoms, higher anxiety scores, and were less likely to consult a doctor despite having CAD symptoms out of fear of infection. Our results provide evidence that the majority of patients received sufficient medical care during the COVID-19 pandemic in Germany. However, one in ten patients could be considered particularly at risk for medical undersupply and adverse health outcomes. The perceived infection risk with COVID-19 might have facilitated the decision not to consult a medical doctor.

Background: Post COVID-19 Condition (PCC, also known as long COVID and post-acute sequelae of COVID-19) is a major public health concern with severe and pervasive impacts on physical and mental health. PCC is highly heterogeneous and may manifest as different clusters of symptoms of varying intensity and duration. The etiology of PCC remains uncertain, though several underlying pathophysiological mechanisms, such as cellular damage, inflammatory cytokines, and a hypercoagulable state, are thought to contribute to PCC inception and trajectory. Examination of potential serological markers of PCC, accounting for clinical covariates, may yield emergent pathophysiological insights. Objectives: Primary objectives of this thesis are to 1) Identify key clinical and potential serological predictors of PCC; 2) Acquire clinical and serological data in a large-scale prospective observational study; 3) Assess relationships between PCC and serological markers, accounting for clinical covariates; 4) Systematically review evidence to date on primary observational studies comparing serological response between people with and without persistent symptoms post COVID-19 recovery; 5) Discuss persisting gaps in knowledge and data quality, and propose strategies for resolve. Methods: This thesis is framed around three core efforts: 1) The design of survey questions and study materials, recruitment of participants, and data collection in a large-scale prospective cohort study launched in 2020; 2) The assessment of relationships between pre-defined serological predictors and PCC, accounting for clinical covariates; and 3) A robust rapid review of PCC onset and phenotype as functions of serological markers. Expert opinion was sought to define serological predictors. Clinical predictors were defined a priori based on systematic reviews meeting AMSTAR 2 guidelines. Conclusions: To address objectives, we described efforts to collect clinical and serological data from a large-scale prospective cohort study; identify PCC-cases and infected-controls; assess associations between pre-defined serological predictors (IgG titres targeting SARS-CoV-2 spike (S), nucleocapsid (N), and receiver binding domain (RBD) antigens, and efficient neutralization) and PCC; and synthesized findings from an extensive rapid review on PCC as a function of serological markers. Our multivariate analysis using Stop the Spread Ottawa data is, to our knowledge, the first Canadian study to report the direction and magnitude of association between selected serological predictors (anti-IgG response to S, N, and RBD SARS-CoV-2 antigens, and neutralizing efficiency) and PCC status and impact on quality of life. Finally, we described five potential strategies which may improve the accessibility, quality, and amalgamation of data pertaining to PCC: 1) Fostering comparability between studies to enable synthesis of multiple datasets; 2) Advancing the characterization and consensus on PCC phenotypes; 3) Employing innovative modelling strategies that could potentially yield novel insights; 4) Promoting robust collaboration and knowledge sharing among research teams; and 5) Engaging people with lived experience at all stages of research.

No description available.

Tesis por compendio / [ES] El objetivo inicial de esta tesis era detectar y rastrear virus entéricos en diferentes matrices acuáticas mediante la combinación de protocolos moleculares y metagenómicos. Se establecieron como objetivos principales el desarrollo de procedimientos para la concentración de virus en muestras de aguas residuales, el análisis de virus indicadores de contaminación fecal y la caracterización del viroma de estas muestras. En cuanto al desarrollo de procedimientos para la concentración de virus entéricos emergentes en muestras de aguas residuales, el protocolo de floculación con hidróxido de aluminio, utilizado habitualmente en el grupo para la concentración de otros virus entéricos, resultó muy eficaz para el virus de la Hepatitis E, permitiendo su detección en muestras de aguas de entrada y salida de estaciones depuradoras de aguas residuales (EDAR). Por lo que respecta al seguimiento de la prevalencia de otros virus entéricos e indicadores virales en muestras de entrada y salida de EDARs, esta tesis proporciona información cuantitativa sobre la presencia del indicador crAssphage y otros virus entéricos de cápside intacta en aguas de diferentes EDARs valencianas. Además, los resultados de la correlación indican que crAssphage podría no ser un indicador óptimo de la presencia de virus entéricos infecciosos en las aguas residuales tratadas. En relación a la caracterización del viroma de las muestras de agua de las EDARs analizadas, en la presente tesis se describe un procedimiento de referencia que permite la detección y caracterización de las poblaciones virales en las muestras de aguas residuales recogidas a la entrada y salida de la planta depuradora. También se refleja el sesgo existente en los perfiles del viroma que se obtienen según las librerías de secuenciación que se empleen. En este sentido, esta investigación arroja luz sobre la diversidad de las comunidades virales en influentes y efluentes de aguas residuales, proporcionando información valiosa también en términos de indicadores fecales virales. Con la llegada de la pandemia de COVID-19 a principios de 2020, el SARS-CoV-pasó a ser el protagonista de la segunda parte de la tesis. En este aspecto, se marcaron como objetivos implementar un sistema de monitorización de SARS-CoV-2 en aguas residuales y desarrollar y optimizar métodos moleculares rápidos para inferir la infectividad del SARS-CoV-2. Los resultados obtenidos han demostrado que la aplicación de la epidemiología basada en aguas residuales (WBE) es eficiente para estimar la presencia de COVID-19 en comunidades y puede servir de herramienta para la salud pública como alerta temprana ante situaciones pandémicas. Asimismo, esta tesis incluye el primer estudio publicado en España que realizó un análisis metagenómico de la diversidad del SARS-CoV-2 presente en las aguas residuales en las tres primeras oleadas epidemiológicas, cuyos resultados confirmaron el potencial de la secuenciación masiva de aguas residuales para detectar nuevas mutaciones y linajes del SARS-CoV-2. Además, también se han comparado y optimizado los protocolos de concentración, extracción y detección de ácidos nucleicos de coronavirus a partir de muestras de aguas residuales, superficiales y de mar. Así, este trabajo amplía el conocimiento sobre los procedimientos analíticos para la detección del SARS-CoV-2 en aguas residuales favoreciendo la implementación global del COVID-19 WBE. Finalmente, se ha implementado un protocolo de RT-qPCR de viabilidad basado en el cloruro de platino para evitar la amplificación del ARN del SARS-CoV-2 no infeccioso. Además, los resultados de esta tesis apoyan la idea de que el SARS-CoV-2 presente en las aguas residuales no es infeccioso. En general, en el marco de esta tesis doctoral se ha desarrollado una herramienta analítica rápida basada en la RT-qPCR de viabilidad para inferir la infectividad del SARS-CoV-2 con potencial aplicación en la evaluación de riesgos, la prevención y el control en los programas de salud / [CA] L'objectiu inicial d'esta tesi era detectar i rastrejar virus entèrics en diferents matrius aquàtiques mitjançant la combinació de protocols moleculars i metagenòmics. Es van establir com a objectius específics el desenvolupament de procediments per a la concentració de virus en mostres d'aigües residuals, l'anàlisi de virus indicadors de contaminació fecal i la caracterització del viroma d'estes mostres. Pel que fa al desenvolupament de procediments per a la concentració de virus entèrics emergents en mostres d'aigües residuals, el protocol de floculació amb hidròxid d'alumini, utilitzat habitualment al grup per a la concentració d'altres virus entèrics, va resultar molt eficaç per al virus de l'hepatitis E, permetent-ne la detecció en mostres d'aigües d'entrada i de sortida d'estacions depuradores d'aigües residuals (EDAR). Pel que fa al seguiment de la prevalença d'altres virus entèrics i indicadors virals en mostres d'entrada i de sortida d'EDARs, esta tesi proporciona informació quantitativa sobre la presència de l'indicador crAssphage i altres virus entèrics de càpside intacta en aigües de diferents EDARs valencianes. A més, els resultats de la correlació indiquen que el bacteriòfag crAssphage podria no ser un indicador òptim de la presència de virus entèrics infecciosos a les aigües residuals tractades. Pel que fa a la caracterització del viroma de les mostres d'aigua de les EDARs analitzades, a la present tesi es descriu un procediment de referència que permet la detecció i caracterització de les poblacions virals a les mostres d'aigües residuals recollides a l'entrada i sortida de la planta depuradora. També es reflecteix el biaix existent als perfils del viroma que s'obtenen segons les llibreries de seqüenciació que s'utilitzen. En este sentit, esta investigació aporta coneixement sobre la diversitat de les comunitats virals en influents i efluents d'aigües residuals, proporcionant informació valuosa també en termes d'indicadors fecals virals. Amb l'arribada de la pandèmia de COVID-19 a principis del 2020, el SARS-CoV-va passar a ser el protagonista de la segona part de la tesi. En este aspecte, es van establir com a objectius implementar un sistema de monitorització de SARS-CoV-2 en aigües residuals i desenvolupar i optimitzar mètodes moleculars ràpids per inferir la infectivitat del SARS-CoV-2. Els resultats obtinguts han demostrat que l'aplicació de l'epidemiologia basada en aigües residuals (WBE) és eficient per estimar la presència de COVID-19 a les comunitats i pot servir d'eina per a la salut pública com a alerta primerenca davant de situacions pandèmiques. Així mateix, esta tesi inclou el primer estudi publicat a Espanya que va realitzar una anàlisi metagenòmica de la diversitat del SARS-CoV-2 present a les aigües residuals a les tres primeres onades epidemiològiques, els resultats del qual van confirmar el potencial de la seqüenciació massiva d'aigües residuals per a detectar noves mutacions i llinatges del SARS-CoV-2. A més, també s'han comparat i optimitzat els protocols de concentració, extracció i detecció d'àcids nucleics de coronavirus a partir de mostres d'aigües residuals, superficials i de mar. Així, este treball amplia el coneixement sobre els procediments analítics per a la detecció del SARS-CoV-2 en aigües residuals afavorint la implementació global del COVID-19 WBE. Finalment, s'ha implementat un protocol de RT-qPCR de viabilitat basat en el clorur de platí per evitar l'amplificació de l'ARN del SARS-CoV-2 no infecciós. A més, els resultats d'esta tesi donen suport a la idea que el SARS-CoV-2 present a les aigües residuals no és infecciós. En general, en el marc d'esta tesi doctoral s'ha desenvolupat una eina analítica ràpida basada en la RT-qPCR de viabilitat per inferir la infectivitat del SARS-CoV-2 amb potencial aplicació en l'avaluació de riscos, la prevenció i el control en els programes de Salut Pública. / [EN] The initial aim of this thesis was to detect and monitor the presence of enteric viruses in different aquatic matrices using both molecular and metagenomic protocols. The specific objectives were the development of procedures for the concentration of viruses in wastewater samples, the analysis of viruses indicative of faecal contamination and the characterisation of the virome in these samples. Regarding the development of procedures for the concentration of emerging enteric viruses in wastewater samples, the flocculation protocol with aluminium hydroxide, commonly used in the group for the concentration of other enteric viruses, proved to be very effective for Hepatitis E virus, allowing its detection in influent and effluent water samples from wastewater treatment plants (WWTPs). Regarding the monitoring of the prevalence of other enteric viruses and viral indicators in incoming and outgoing samples from WWTPs, this thesis provides quantitative information on the presence of the indicator crAssphage and other enteric viruses with intact capsid in water from different Valencian WWTPs. Moreover, the correlation results indicate that crAssphage may not be an optimal indicator of the presence of infectious enteric viruses in treated wastewater. In relation to the virome characterisation of the WWTP water samples analysed, the present thesis describes a reference procedure that allows the detection and characterisation of viral populations in wastewater. It also reflects the existing bias in the virome profiles obtained depending on the sequencing libraries used. In this sense, this research sheds light on the diversity of viral communities in influent and effluent wastewater, providing valuable information also in terms of faecal viral indicators. With the advent of the COVID-19 pandemic in the early 2020s, SARS-CoV-2 became the focus of the second part of the thesis. In this aspect, the objectives were to implement a monitoring system for SARS-CoV-2 in wastewater and to develop and optimise rapid molecular methods to infer SARS-CoV-2 infectivity. The results obtained have demonstrated that the application of wastewater-based epidemiology (WBE) is efficient for estimating the presence of COVID-19 in communities and can serve as a Public Health tool for early warning of pandemic situations. Furthermore, this thesis includes the first study published in Spain that performed a metagenomic analysis of the SARS-CoV-2 diversity present in wastewater in the first three epidemiological waves, the results of which confirmed the potential of mass sequencing of wastewater to detect new SARS-CoV-2 mutations and lineages. In addition, protocols for the concentration, extraction and detection of coronavirus nucleic acids from sewage, surface and seawater samples have also been compared and optimised. Thus, this work expands the knowledge on analytical procedures for the detection of SARS-CoV-2 in wastewater favouring the global implementation of the COVID-19 WBE. Finally, a viability RT-qPCR protocol based on platinum chloride has been implemented to avoid amplification of non-infectious SARS-CoV-2 RNA. Furthermore, the results of this thesis support the idea that SARS-CoV-2 present in wastewater is not infectious. Overall, in the framework of this PhD thesis, a rapid analytical tool based on feasibility RT-qPCR has been developed to infer the infectivity of SARS-CoV-2 with potential application in risk assessment, prevention and control in Public Health programmes. / This thesis has been funded by an "Ayudas para contratos predoctorales para la formación de doctores 2018” grant, awarded by the Ministry of Science and Innovation. / Cuevas Ferrando, E. (2022). Detection and tracking of emerging viruses of public health interest in waters through molecular and metagenomic procedures [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/190899 / Compendio

Coronavirus eller SARS-CoV-2 (severe acute respiratory syndrom coronavirus 2) som fick utbrott i december 2019 har väckt en stor uppmärksamhet och påverkat mänskligheten världen runt och inte minst påverkat den socioekonomiska balansen. Viruset som har ursprung i staden Wuhan i Kina sprider sig snabbt mellan människorna genom frekvent rekombination av det genetiska materialet. Redan efter ett år från utbrottet beräknades antal fall till 98 miljoner och dödsfallen till 2 miljoner globalt. Förutom lunginflammation leder infektionen till högre halter av proinflammatoriska markörer som CRP och höga nivåer av cytokiner som IL-6 som i slutändan kan resultera i en cytokinstorm. Därför anses en blockering av IL6 produktionen och/eller blockering av receptorbindningen vara en terapeutisk lösning för att begränsa patogenicitet. Tocilizumab som är den första immunmodulator som introducerades och testades mot covid-19 pneumoni har i flera studier gett upphov till heterogent resultat angående dess effekt på sjukdomen. Syftet med detta litteraturabete är att studera tocilizumabs effekt på covid-19 sjuka patienter med pneumoni och med måttlig till svår sjukdom. Genom sökning på databasen PubMed hittades sex randomiserade kliniska studier som valdes för att undersökas i detta arbete. Studierna jämförde effekten av standardbehandling med och utan tocilizumab. Resultatet blev heterogent men en signifikant förbättring observerades för tocilizumab vad gäller överlevnad, sjukdomsprogression och hälsostatus. Trots att tocilizumabs fördelar överväger dess nackdelar krävs det dock mer forskning kring läkemedlet för att kunna dra bättre samband mellan tocilizumab och dess effekter.

Importance: Immunological response to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccination is important, especially in people with multiple sclerosis (pwMS) on immunosuppressive therapies. Objective: This study aims to determine whether adjuvanted protein-based vaccine NVX-CoV2373 is able to induce an immune response to SARS-CoV-2 in pwMS with inadequate responses to prior triple mRNA/viral vector vaccination. Design, setting, and participants: We conducted a single-center, prospective longitudinal cohort study at the MS Center in Dresden, Germany. In total, 65 participants were included in the study in accordance with the following eligibility criteria: age > 18 years, immunomodulatory treatment, and insufficient T-cellular and humoral response to prior vaccination with at least two doses of SARS-CoV-2 mRNA (BNT162b2, mRNA-1273) or viral vector vaccines (AZD1222, Ad26.COV2.S). Interventions: Intramuscular vaccination with two doses of NVX-CoV2373 at baseline and 3 weeks of follow-up. Main outcomes and measures: The development of SARS-CoV-2-specific antibodies and T-cell responses was evaluated. Results: For the final analysis, data from 47 patients on stable treatment with sphingosine-1-phosphate receptor (S1PR) modulators and 17 on ocrelizumab were available. The tolerability of the NVX-CoV2373 vaccination was overall good and comparable to the one reported for the general population. After the second NVX-CoV2373 vaccination, 59% of S1PR-modulated patients developed antispike IgG antibodies above the predefined cutoff of 200 binding antibody units (BAU)/ml (mean, 1,204.37 [95% CI, 693.15, 2,092.65] BAU/ml), whereas no clinically significant T-cell response was found. In the subgroup of the patients on ocrelizumab treatment, 23.5% developed antispike IgG > 200 BAU/ml (mean, 116.3 [95% CI, 47.04, 287.51] BAU/ml) and 53% showed positive spike-specific T-cellular responses (IFN-gamma release to antigen 1: mean, 0.2 [95% CI, 0.11, 0.31] IU/ml; antigen 2: mean, 0.24 [95% CI, 0.14, 0.37]) after the second vaccination. Conclusions: Vaccination with two doses of NVX-CoV2373 was able to elicit a SARS-CoV-2-specific immune response in pwMS lacking adequate immune responses to previous mRNA/viral vector vaccination. For patients receiving S1PR modulators, an increase in anti-SARS-CoV-2 IgG antibodies was detected after NVX-CoV2373 vaccination, whereas in ocrelizumab-treated patients, the increase of antiviral T-cell responses was more pronounced. Our data may impact clinical decision-making by influencing the preference for NVX-CoV2373 vaccination in pwMS receiving treatment with S1PR modulation or anti-CD20 treatment.

COVID-19 pandemic has exposed both strengths and weaknesses in world communities and particularly put a severe strain on health care. It is widely accepted that lifestyle factors are an important piece of the puzzle in preventive work against infectious diseases. One of these lifestyle factors is physical activity (FA) that positively affects people's well-being and has a strengthening effect on, among other things, the immune system. The purpose of the study was to investigate how people that have had SARS-CoV-2 infection describe their FA level and general state of health before the illness and how they feel that it has affected their disease course. A total of 10 semi-structured interviews were conducted during the period FebruaryMarch, which were then interpreted using content analysis as a tool. During the analysis work three main categories were identified: Well-balanced FA level, Relatively good general health, Experience of COVID-19 and FA significance. Most respondents perceived themselves as physically active to a greater or lesser extent. Respondents' perception indicates that FA has an important role in facing COVID-19 disease. Results indicate an increasingly accepted view in research that FA can act as a protective factor against SARS-CoV-2 virus / <p>Betyg i Ladok 220613.</p>

京都大学 / 新制・課程博士 / 博士(医科学) / 甲第25202号 / 医科博第158号 / 新制||医科||10(附属図書館) / 京都大学大学院医学研究科医科学専攻 / (主査)教授 川口 義弥, 教授 波多野 悦朗, 教授 妹尾 浩 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DFAM