184 research outputs found

    Quantum virology : improved management of viral infections through quantitative measurements

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    Real-time monitoring of PCR has strongly supported the increased diagnostic use of nucleic acid detection assays in clinical virology. Particularly the improvements in the ability to quantify target nucleic acid sequences offer new opportunities in the management of viral infections. Real-time PCR is rapidly replacing traditional PCR, and new diagnostic uses will likely emerge. This thesis explores the wide range of potential applications of real-time quantitative PCR technology in clinical virology. This exploration is directed to the design of methods, the application to relevant patient categories, the comparison with established methods where available, and the definition of the clinical relevance of the approach. The focus comprises viral targets where an elaborate balance between viral replication and the host immune system has been established, which brings about viral maintenance without affecting the host, until this balance is disturbed.LEI Universiteit LeidenMoleculaire basis van virus pathogenese en antivirale strategiee

    Satellite radar observation feasibility for large infrastructure public works

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    A very large infrastructural work is being undertaken in Delft. A tunnel is going to be constructed to replace the current rail viaduct. As in any large infrastructural work, the monitoring of the land deformations during the period of the tunnel’s construction is highly essential. In this project, one study was performed to analyse the feasibility of PSInSAR as an independent technique for monitoring of land subsidence. The driving mechanisms for deformation were studied to find out the relations among them. Soil geophysics, hydrology, infrastructures and thermal expansion were studied in relation with deformation. Besides these, the traditional deformation monitoring methods were also studied, since they are the competitors of this technique. LiDAR, photogrammetry, tachymetry, levelling and GPS were considered in this study for the comparison with radar. The major attention has been given in this study to assessment of the geolocalisation quality of PSInSAR observations. We have found that the accuracy and point density of PSInSAR is sufficient for deformation monitoring. While its repeat interval might not be able to detect quick failure mechanisms, other techniques have proven to be excellent complements for this deficiency. Radar measurements are also good for validation in other fields, showing that they correlate well with thermal expansion and soil mechanics theory. Further work should be directed to improving geolocalisation and deformation models

    Assessment of Cytomegalovirus-Specific Cell-Mediated Immunity for the Prediction of Cytomegalovirus Disease in High-Risk Solid-Organ Transplant Recipients: A Multicenter Cohort Study

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    In high-risk solid-organ transplant recipients receiving antiviral prophylaxis, the measurement of specific cell-mediated immunity using the Quantiferon assay appropriately stratified the individual risk of developing subsequent cytomegalovirus diseas

    Post COVID-19 Pandemic Increased Detection of Mycoplasma Pneumoniae in Adults Admitted to the Intensive Care

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    Background: Mycoplasma pneumoniae (M. pneumoniae) infections can progress to severe respiratory complications, necessitating intensive care treatment. Recent post COVID-19 pandemic surges underscore the need for timely diagnosis, given potential diagnostic method limitations. Methods: A retrospective case series analysis was conducted on M. pneumonia PCR-positive patients admitted to two Dutch secondary hospitals’ ICUs between January 2023 and February 2024. Clinical presentations, treatments, outcomes, and mechanical ventilation data were assessed. Results: Seventeen ICU-admitted patients were identified, with a median age of 44 years, primarily due to hypoxia. Non-invasive ventilation was effective for most, while five required invasive mechanical ventilation. None of the patients required extracorporeal membrane oxygenation. No fatalities occurred. Post-PCR, treatment was adjusted to doxycycline or azithromycin; seven received steroid treatment. Discussion: Increased ICU admissions for M. pneumoniae infection were observed. Diverse clinical and radiological findings emphasize heightened clinical awareness. Early molecular diagnostics and tailored antibiotic regimens are crucial since beta-lactam antibiotics are ineffective. Conclusion: This study highlights the escalating challenge of severe M. pneumoniae infections in ICUs, necessitating a multifaceted approach involving accurate diagnostics, vigilant monitoring, and adaptable treatment strategies for optimal patient outcomes.</p

    Comparison of Quantitative Cytomegalovirus Real-time PCR in Whole Blood and pp65 Antigenemia Assay: Clinical Utility of CMV Real-time PCR in Hematopoietic Stem Cell Transplant Recipients

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    Successful preemptive therapy for cytomegalovirus (CMV) infection in transplant patients depends on the availability of sensitive, specific, and timely diagnostic tests for CMV infection. Although the pp65 antigenemia assay has been widely used for this purpose, real-time quantification of CMV DNA has recently been recognized as an alternative diagnostic approach. However, the guidelines for antiviral therapy based on real-time quantitative polymerase chain reaction (RQ-PCR) have yet to be established. From November 2004 to March 2005, a total of 555 whole blood samples from 131 hematopoietic stem cell transplant (HSCT) recipients were prospectively collected. RQ-PCR was conducted using an Artus® CMV LC PCR kit (QIAGEN). Both qualitative and quantitative correlations were drawn between the two methods. Exposure to the antiviral agent influenced the results of the two assays. Additionally, the discrepancy was observed at low levels of antigenemia and CMV DNA load. Via ROC curve analysis, the tentative cutoff value for preemptive therapy was determined to be approximately 2×104 copies/mL (sensitivity, 80.0%; specificity, 50.0%) in the high risk patients, and approximately 3×104 copies/mL (sensitivity, 90.0%; specificity, 70.0%) in the patients at low risk for CMV disease. Further study to validate the optimal cutoff value for the initiation of preemptive therapy is currently underway

    Relationship between pp65 antigenemia levels and real-time quantitative DNA PCR for Human Cytomegalovirus (HCMV) management in immunocompromised patients

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    <p>Abstract</p> <p>Background</p> <p>Quantitative real-time PCR assays, which are more rapid and practical than pp65 antigenemia determination, are progressively becoming the preferred method for monitoring Human Cytomegalovirus (HCMV) reactivation. However, the relationship between HCMV DNA and antigenemia levels is still under investigation. The aim of this study was to analyse the relationship between HCMV DNA and pp65 antigenemia levels in order to identify clinically useful threshold values for the management of patients.</p> <p>Methods</p> <p>475 consecutive samples from 156 immunosuppressed patients were tested for HCMV by pp65 antigenemia and Real-time PCR assay.</p> <p>Results </p> <p>136 out of 475 consecutive samples derived from 48 patients showed evidence of HCMV infection. HCMV DNA was detected in 106 samples, pp65 antigen in 3, and both markers in 27. pp65 antigen detection was associated with higher HCMV DNA levels. The cut-off HCMV DNA level that best predicted pp65 antigenemia in this series of samples was 11,500 copies/ml, but different threshold levels could be observed for specific groups of patients. HCMV disease was observed in 5 out of 48 patients with active HCMV infection. The presence of clinical symptoms was associated with positive pp65 and with higher antigenemia levels. Higher HCMV DNA load at the onset of viral replication was correlated to the development of clinical symptoms.</p> <p>Conclusion</p> <p>Both pp65 antigenemia and HCMV DNA load can be useful for the prospective monitoring of immunocompromised subjects. Specific cut-off levels capable of triggering preemptive antiviral treatment should be determined in accordance to the type of test used and the characteristics of patients and prospectively validated.</p
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