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Background & Aims Currently available HDV PCR assays are characterized by considerable run-to-run and inter-laboratory variability. Hence, we established a quantitative RT-PCR assay on the open channel of a fully automated PCR platform (cobas6800, Roche) offering improved consistency and reliability. Methods A primer/probe-set targeting a highly conserved region upstream of the HDV-antigen was adapted for use on the cobas6800. The lower limit of detection (LOD) was determined using a dilution panel of the HDV WHO standard (n=21/dilution). Linearity and inclusivity were tested by preparing 10-fold dilution series (genotypes (GT) 1-8 cell culture-derived; n=5/dilution). Patient samples containing a variety of bloodborne viral pathogens were tested to confirm exclusivity (n=60). Results LOD of the HDV Utility-Channel assay (HDV_UTC) was determined as 3.86 IU/ml (95%-CI:2.95-5.05 IU/ml) with a linear range from 10-10ˆ8 IU/ml (GT1). Linear relationships were observed for all HDV GT with slopes ranging from -3.481 to -4.134 cycles/log and R2 from 0.918 to 0.994. Inter-run and intra-run variability was 0.3 and 0.6 Ct (3xLOD), respectively. No false-positive results were observed. To evaluate clinical performance, 110 serum samples of anti-HDV-Ab+ patients were analyzed using the HDV_UTC and CE-IVD RoboGene assay. 58/110 and 49/110 samples were concordant positive or negative, respectively (overall agreement 97,3%). Quantitative comparison demonstrated a strong correlation (r:0.922; 95%-CI:0.869-0.954; p-value:<0.0001). Conclusion The use of highly automated, sample-to-result solutions for molecular diagnostics holds many inherent benefits over manual workflows, including improved reliability, reproducibility and dynamic scaling of testing capacity. The assay we established showed excellent analytical and clinical performance, with inclusivity for all HDV GT and a limit of quantification of 10 IU/ml, making it a sensitive new tool for HDV screening and viral load monitoring. Lay Summary The hepatitis delta virus (HDV) causes a severe form of inflammation in the liver. We developed a tool for molecular diagnostics, a polymerase chain reaction HDV assay that showed great performance. It can be used to improve diagnosis of HDV, as well as for monitoring of treatment success. The assay allows for quantification of the virus in the tested samples and is performed on a fully automated platform (cobas6800), which provides various benefits including less hands-on time and excellent comparability of test results.
Lisa Sophie Pflüger; Dominik Nörz; Tassilo Volz; Katja Giersch; Annika Giese; Nora Goldmann; Dieter Glebe; Jan-Hendrik Bockmann; Susanne Pfefferle; Maura Dandri; Julian Schulze Zur Wiesch; Marc Lütgehetmann. Clinical establishment of a laboratory developed quantitative HDV PCR assay on the cobas6800 high-throughput system. JHEP Reports 2021, 1 .
AMA StyleLisa Sophie Pflüger, Dominik Nörz, Tassilo Volz, Katja Giersch, Annika Giese, Nora Goldmann, Dieter Glebe, Jan-Hendrik Bockmann, Susanne Pfefferle, Maura Dandri, Julian Schulze Zur Wiesch, Marc Lütgehetmann. Clinical establishment of a laboratory developed quantitative HDV PCR assay on the cobas6800 high-throughput system. JHEP Reports. 2021; ():1.
Chicago/Turabian StyleLisa Sophie Pflüger; Dominik Nörz; Tassilo Volz; Katja Giersch; Annika Giese; Nora Goldmann; Dieter Glebe; Jan-Hendrik Bockmann; Susanne Pfefferle; Maura Dandri; Julian Schulze Zur Wiesch; Marc Lütgehetmann. 2021. "Clinical establishment of a laboratory developed quantitative HDV PCR assay on the cobas6800 high-throughput system." JHEP Reports , no. : 1.
Molecular testing for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to suffer from delays and shortages. Antigen tests have recently emerged as a viable alternative to detect patients with high viral loads, associated with elevated risk of transmission. While rapid lateral flow tests greatly improved accessibility of SARS-CoV-2 detection in critical areas, their manual nature limits scalability and suitability for large-scale testing schemes. The Elecsys® SARS-CoV-2 Antigen assay allows antigen immunoassays to be carried out on fully automated high-throughput serology platforms. A total of 3139 nasopharyngeal and oropharyngeal swabs were collected at 3 different testing sites in Germany. Swab samples were pre-characterized by reverse transcription real-time polymerase chain reaction (RT-qPCR) and consecutively subjected to the antigen immunoassay on either the cobas e 411 or cobas e 801 analyzer. Of the tested respiratory samples, 392 were PCR positive for SARS-CoV-2 RNA. Median concentration was 2.95 × 104 (interquartile range [IQR] 5.1 × 102–3.5 × 106) copies/ml. Overall sensitivity and specificity of the antigen immunoassay were 60.2% (95% confidence interval [CI] 55.2–65.1) and 99.9% (95% CI 99.6–100.0), respectively. A 93.7% (95% CI 89.7–96.5) sensitivity was achieved at a viral RNA concentration ≥ 104 copies/ml (~ cycle threshold [Ct] value < 29.9). The Elecsys SARS-CoV-2 Antigen assay reliably detected patient samples with viral loads ≥ 10,000 copies/ml. It thus represents a viable high-throughput alternative for screening of patients or in situations where PCR testing is not readily available. The online version contains supplementary material available at 10.1007/s40121-021-00510-x.
Dominik Nörz; Flaminia Olearo; Stojan Perisic; Matthias F. Bauer; Elena Riester; Tanja Schneider; Kathrin Schönfeld; Tina Laengin; Marc Lütgehetmann. Multicenter Evaluation of a Fully Automated High-Throughput SARS-CoV-2 Antigen Immunoassay. Infectious Diseases and Therapy 2021, 1 -9.
AMA StyleDominik Nörz, Flaminia Olearo, Stojan Perisic, Matthias F. Bauer, Elena Riester, Tanja Schneider, Kathrin Schönfeld, Tina Laengin, Marc Lütgehetmann. Multicenter Evaluation of a Fully Automated High-Throughput SARS-CoV-2 Antigen Immunoassay. Infectious Diseases and Therapy. 2021; ():1-9.
Chicago/Turabian StyleDominik Nörz; Flaminia Olearo; Stojan Perisic; Matthias F. Bauer; Elena Riester; Tanja Schneider; Kathrin Schönfeld; Tina Laengin; Marc Lütgehetmann. 2021. "Multicenter Evaluation of a Fully Automated High-Throughput SARS-CoV-2 Antigen Immunoassay." Infectious Diseases and Therapy , no. : 1-9.
Few data are available regarding the efficacy of anti-SARS-CoV-2 vaccines in patients with hematological malignancies, and particular, plasma cell neoplasia. This ongoing single-center study aimed to describe the level of post-vaccination anti-SARS-CoV-2-antibodies depending on B lymphocyte count, current therapy, and remission status of patients with multiple myeloma and related plasma cell dyscrasia, after the first dose of anti-SARS-CoV-2 vaccination. The 82 patients included in this study received SARS-CoV-2 vaccines (including mRNA- and vector-based vaccines) as a routine measure. After the first vaccination, a positive SARS-CoV-2 spike protein antibody titer (SP-AbT) was detected in 23% of assessable patients. SARS-CoV-2 SP-AbT was significantly higher in patients with higher CD19+ B lymphocyte counts. A cut-off value of ≥30 CD19+ B cells/µL was significantly positive correlating with higher SARS-CoV-2 SP-AbT. In contrast, current treatment with anti-CD38-antibodies has led to significantly reduced SP-AbT titers. Furthermore, in multivariable linear regression, higher age and insufficiently controlled disease significantly correlated negatively with SARS-CoV-2 SP-AbT. Conversely, treatment with immunomodulatory drugs did not harm the development of antibody titers. Based on our results, the majority of myeloma patients respond poorly after receiving the first dose of any anti-SARS-CoV-2 vaccination and need booster vaccination.
Susanne Ghandili; Martin Schönlein; Marc Lütgehetmann; Julian Schulze Zur Wiesch; Heiko Becher; Carsten Bokemeyer; Marianne Sinn; Katja Weisel; Lisa Leypoldt. Post-Vaccination Anti-SARS-CoV-2-Antibody Response in Patients with Multiple Myeloma Correlates with Low CD19+ B-Lymphocyte Count and Anti-CD38 Treatment. Cancers 2021, 13, 3800 .
AMA StyleSusanne Ghandili, Martin Schönlein, Marc Lütgehetmann, Julian Schulze Zur Wiesch, Heiko Becher, Carsten Bokemeyer, Marianne Sinn, Katja Weisel, Lisa Leypoldt. Post-Vaccination Anti-SARS-CoV-2-Antibody Response in Patients with Multiple Myeloma Correlates with Low CD19+ B-Lymphocyte Count and Anti-CD38 Treatment. Cancers. 2021; 13 (15):3800.
Chicago/Turabian StyleSusanne Ghandili; Martin Schönlein; Marc Lütgehetmann; Julian Schulze Zur Wiesch; Heiko Becher; Carsten Bokemeyer; Marianne Sinn; Katja Weisel; Lisa Leypoldt. 2021. "Post-Vaccination Anti-SARS-CoV-2-Antibody Response in Patients with Multiple Myeloma Correlates with Low CD19+ B-Lymphocyte Count and Anti-CD38 Treatment." Cancers 13, no. 15: 3800.
Carbapenem-resistant Gram-negative bacteria (CR-GNB) are a major source of nosocomial infections worldwide. In this study, the ability of a loop-mediated isothermal amplification (LAMP)-based method (Isoplex CRE-ART) to rapidly detect carbapenemase-encoding genes bla OXA-48-like, bla OXA-23-like, bla OXA-24-like, bla KPC, bla VIM, bla NDM and bla IMP in 231 carbapenem-resistant Enterobacterales, Pseudomonas aeruginosa and Acinetobacter baumannii isolates was investigated. The accuracy of the LAMP test was compared to results of molecular isolate characterization using a Laboratory Developed Test multiplex carbapenemase PCR assay. The LAMP test correctly identified the presence of on-panel carbapenemases with a sensitivity of 99.16 % [95 % confidence interval (CI): 95.39–99.96 %] and a specificity of 98.21 % (95 % CI: 93.72–99.68 %) in 60 min. Our findings suggest that the Isoplex CRE-ART assay is able to rapidly identify carbapenemase genes in CR-GNB and improves options for pathogen characterization in the context of clinical microbiological and infection control diagnostics.
Laura Berneking; Lucia Asar; Anna Both; Benjamin Berinson; Martin Aepfelbacher; Marc Lütgehetmann; Holger Rohde. Performance of a loop-mediated isothermal amplification assay (Isoplex CRE-ART) to detect common carbapenemase-encoding genes in Gram-negative bacteria. Journal of Medical Microbiology 2021, 70, 001379 .
AMA StyleLaura Berneking, Lucia Asar, Anna Both, Benjamin Berinson, Martin Aepfelbacher, Marc Lütgehetmann, Holger Rohde. Performance of a loop-mediated isothermal amplification assay (Isoplex CRE-ART) to detect common carbapenemase-encoding genes in Gram-negative bacteria. Journal of Medical Microbiology. 2021; 70 (7):001379.
Chicago/Turabian StyleLaura Berneking; Lucia Asar; Anna Both; Benjamin Berinson; Martin Aepfelbacher; Marc Lütgehetmann; Holger Rohde. 2021. "Performance of a loop-mediated isothermal amplification assay (Isoplex CRE-ART) to detect common carbapenemase-encoding genes in Gram-negative bacteria." Journal of Medical Microbiology 70, no. 7: 001379.
Critical Coronavirus disease 2019 (COVID-19) developed in a 7-year-old girl with a history of dystrophy, microcephaly, and central hypothyroidism. Starting with gastrointestinal symptoms, the patient developed severe myocarditis followed by progressive multiple organ failure complicated by Pseudomonas aeruginosa bloodstream infection. Intensive care treatment consisting of invasive ventilation, drainage of pleural effusion, and high catecholamine therapy could not prevent the progression of heart failure, leading to the implantation of venoarterial extracorporeal life support (VA-ECLS) and additional left ventricle support catheter (Impella® pump). Continuous venovenous hemofiltration (CVVH) and extracorporeal hemadsorption therapy (CytoSorb®) were initiated. Whole exome sequencing revealed a mutation of unknown significance in DExH-BOX helicase 30 (DHX30), a gene encoding a RNA helicase. COVID-19 specific antiviral and immunomodulatory treatment did not lead to viral clearance or control of hyperinflammation resulting in the patient’s death on extracorporeal life support-(ECLS)-day 20. This fatal case illustrates the potential severity of pediatric COVID-19 and suggests further evaluation of antiviral treatment strategies and vaccination programs for children.
Sofia Apostolidou; Theresa Harbauer; Peter Lasch; Daniel Biermann; Maja Hempel; Marc Lütgehetmann; Susanne Pfefferle; Jochen Herrmann; André Rüffer; Konrad Reinshagen; Rainer Kozlik-Feldmann; Anna Gieras; Inga Kniep; Jun Oh; Dominique Singer; Chinedu Ebenebe; Robin Kobbe. Fatal COVID-19 in a Child with Persistence of SARS-CoV-2 Despite Extensive Multidisciplinary Treatment: A Case Report. Children 2021, 8, 564 .
AMA StyleSofia Apostolidou, Theresa Harbauer, Peter Lasch, Daniel Biermann, Maja Hempel, Marc Lütgehetmann, Susanne Pfefferle, Jochen Herrmann, André Rüffer, Konrad Reinshagen, Rainer Kozlik-Feldmann, Anna Gieras, Inga Kniep, Jun Oh, Dominique Singer, Chinedu Ebenebe, Robin Kobbe. Fatal COVID-19 in a Child with Persistence of SARS-CoV-2 Despite Extensive Multidisciplinary Treatment: A Case Report. Children. 2021; 8 (7):564.
Chicago/Turabian StyleSofia Apostolidou; Theresa Harbauer; Peter Lasch; Daniel Biermann; Maja Hempel; Marc Lütgehetmann; Susanne Pfefferle; Jochen Herrmann; André Rüffer; Konrad Reinshagen; Rainer Kozlik-Feldmann; Anna Gieras; Inga Kniep; Jun Oh; Dominique Singer; Chinedu Ebenebe; Robin Kobbe. 2021. "Fatal COVID-19 in a Child with Persistence of SARS-CoV-2 Despite Extensive Multidisciplinary Treatment: A Case Report." Children 8, no. 7: 564.
New SARS-CoV-2 variants with increased transmissibility, like B.1.1.7, first detected in England or B.1.351, first detected in South Africa, have caused considerable concern worldwide. In order to contain the spread of these lineages, it is of utmost importance to have rapid, sensitive and high-throughput detection methods at hand. A set of RT-qPCR assays was modified for a diagnostic SARS-CoV-2 multiplex assay including detection of the del-HV69/70 and N501Y mutations on the cobas6800 platform. Analytical sensitivity was assessed for both wild-type SARS-CoV-2 and B.1.1.7 lineage by serial dilution. For clinical performance, a total of 176 clinical samples were subjected to the test and results compared to a commercial manual typing-PCR assay and next generation sequencing as gold standard. The multiplex assay was highly sensitive for detection of SARS-CoV-2 RNA in clinical samples, with an LoD of 6.16 cp/ml (CI: 4.00–8.31). LoDs were slightly higher for detection of the HV69/70 deletion (85.92, CI: 61–194.41) and the N501Y SNP (105.99 cp/ml, CI: 81.59 – 183.66). A total of 176 clinical samples were tested with the assay, including 50 samples containing SARS-CoV-2 of the B.1.1.7 lineage, one containing B.1.351 and 85 non-B.1.1.7/B.1.351 lineage, of which three also harbored a HV69/70 deletion. All were correctly identified by the multiplex assay. We describe here a highly sensitive, fully automated multiplex PCR assay for the simultaneous detection of the del-HV69/70 and N501Y mutations that can distinguish between B.1.1.7 and other lineages. The assay allows for high-throughput screening for currently relevant variants in clinical samples prior to sequencing.
Dominik Nörz; Moritz Grunwald; Flaminia Olearo; Nicole Fischer; Martin Aepfelbacher; Susanne Pfefferle; Marc Lütgehetmann. Evaluation of a fully automated high-throughput SARS-CoV-2 multiplex qPCR assay with built-in screening functionality for del-HV69/70- and N501Y variants such as B.1.1.7. Journal of Clinical Virology 2021, 141, 104894 .
AMA StyleDominik Nörz, Moritz Grunwald, Flaminia Olearo, Nicole Fischer, Martin Aepfelbacher, Susanne Pfefferle, Marc Lütgehetmann. Evaluation of a fully automated high-throughput SARS-CoV-2 multiplex qPCR assay with built-in screening functionality for del-HV69/70- and N501Y variants such as B.1.1.7. Journal of Clinical Virology. 2021; 141 ():104894.
Chicago/Turabian StyleDominik Nörz; Moritz Grunwald; Flaminia Olearo; Nicole Fischer; Martin Aepfelbacher; Susanne Pfefferle; Marc Lütgehetmann. 2021. "Evaluation of a fully automated high-throughput SARS-CoV-2 multiplex qPCR assay with built-in screening functionality for del-HV69/70- and N501Y variants such as B.1.1.7." Journal of Clinical Virology 141, no. : 104894.
Background: Neuralgic amyotrophy (NA) has been described as a possible extrahepatic manifestation of hepatitis E virus (HEV) infection. Usually, HEV-associated NA occurs bilaterally. The clinical characteristics determining the course of HEV-associated NA have still not been defined. Methods: In this retrospective multicentric case series, 16 patients with HEV-associated NA were studied and compared to 176 HEV patients without NA in terms of their age, sex, and ALT levels. Results: Neither gender distribution (75% vs. 67% male) nor age (47 vs. 48 years median) differed significantly between the NA patients and controls. Eight NA patients (50%) presented with bilateral involvement—seven of these had right-side dominance and one had left-side dominance. Thirteen cases (81%) were hospitalized. Eight of these patients stayed in hospital for five to seven days, and five patients stayed for up to two weeks. The time from the onset of NA to the HEV diagnosis, as well as the diagnostic and therapeutic proceedings, showed a large variability. In total, 13 (81%) patients received treatment: 1/13 (8%) received intravenous immunoglobulins, 8/13 (62%) received glucocorticoids, 3/13 (23%) received ribavirin, and 6/13 (46%) received pregabalin/gabapentin. Patients with ages above the median (47 years) were more likely to be treated (p = 0.001). Conclusion: HEV-associated NA causes a relevant morbidity. In our case series neither the type of treatment nor the time of initiation of therapy had a significant effect on the duration of hospitalization or the course of the disease. The clinical presentation, the common diagnostic and therapeutic procedures, and the patients’ characteristics showed large variability, demonstrating the necessity of standardized protocols for this rare but relevant disease.
Johannes Bannasch; Benjamin Berger; Claus-Peter Schwartkop; Marco Berning; Oliver Goetze; Marcus Panning; Miriam Fritz-Weltin; George Trendelenburg; Mathias Gelderblom; Marc Lütgehetmann; Fridrike Stute; Thomas Horvatits; Meike Dirks; Christoph Antoni; Patrick Behrendt; Sven Pischke. HEV-Associated Neuralgic Amyotrophy: A Multicentric Case Series. Pathogens 2021, 10, 672 .
AMA StyleJohannes Bannasch, Benjamin Berger, Claus-Peter Schwartkop, Marco Berning, Oliver Goetze, Marcus Panning, Miriam Fritz-Weltin, George Trendelenburg, Mathias Gelderblom, Marc Lütgehetmann, Fridrike Stute, Thomas Horvatits, Meike Dirks, Christoph Antoni, Patrick Behrendt, Sven Pischke. HEV-Associated Neuralgic Amyotrophy: A Multicentric Case Series. Pathogens. 2021; 10 (6):672.
Chicago/Turabian StyleJohannes Bannasch; Benjamin Berger; Claus-Peter Schwartkop; Marco Berning; Oliver Goetze; Marcus Panning; Miriam Fritz-Weltin; George Trendelenburg; Mathias Gelderblom; Marc Lütgehetmann; Fridrike Stute; Thomas Horvatits; Meike Dirks; Christoph Antoni; Patrick Behrendt; Sven Pischke. 2021. "HEV-Associated Neuralgic Amyotrophy: A Multicentric Case Series." Pathogens 10, no. 6: 672.
Introduction Molecular testing for SARS-CoV-2 continues to suffer from delays and shortages. Antigen tests have recently emerged as a viable alternative to detect patients with high viral loads, associated with elevated risk of transmission. While rapid lateral flow tests greatly improved accessibility of SARS-CoV-2 detection in critical areas, their manual nature limits scalability and suitability for large-scale testing schemes. The Elecsys® SARS-CoV-2 Antigen assay allows antigen immunoassays to be carried out on fully automated high-throughput serology platforms. Methods A total of 3139 nasopharyngeal and oropharyngeal swabs were collected at 3 different testing sites in Germany. Swab samples were pre-characterized by RT-qPCR and consecutively subjected to the antigen immunoassay on either the cobas e 411 or cobas e 801 analyzers. Results Of the tested respiratory samples, 392 were PCR positive for SARS-CoV-2 RNA. Median concentration was 2.95×104 (interquartile range [IQR] 5.1×102–3.5×106) copies/mL. Overall sensitivity and specificity of the antigen immunoassay were 60.2% (95% confidence interval [CI] 55.2–65.1) and 99.9% (95% CI 99.6–100), respectively. A 93.7% (95% CI 89.7–96.5) sensitivity was achieved at a viral RNA concentration ≥104 copies/mL (∼cycle threshold (Ct) value<29.9). Conclusion The Elecsys SARS-CoV-2 Antigen assay reliably detected patient samples with viral loads of 10,000 copies/mL and higher. It thus represents a viable high-throughput alternative for screening of patients, or in situations where PCR testing is not readily available. Key Summary Points Why carry out this study? The SARS-CoV-2 pandemic has led to a surge in demand for reliable, mass diagnostic tests worldwide. A thorough clinical evaluation of a fully automated high-throughput Elecsys® SARS-CoV-2 Antigen assay on a total of 3139 clinical samples pre-characterized by quantitative RT-PCR was carried out. What was learned from the study? The assay demonstrated excellent specificity (99.9%) and good relative sensitivity, with an overall sensitivity of 60.2% and a sensitivity of 93.7% for samples containing ≥104 viral RNA copies/mL. The Elecsys SARS-CoV-2 Antigen assay is a viable high-throughput, automated alternative to manual lateral-flow antigen tests.
Dominik Nörz; Flaminia Olearo; Stojan Perisic; Matthias F. Bauer; Elena Riester; Tanja Schneider; Kathrin Schönfeld; Tina Laengin; Marc Lütgehetmann. Multicenter evaluation of a fully automated high-throughput SARS-CoV-2 antigen immunoassay. 2021, 1 .
AMA StyleDominik Nörz, Flaminia Olearo, Stojan Perisic, Matthias F. Bauer, Elena Riester, Tanja Schneider, Kathrin Schönfeld, Tina Laengin, Marc Lütgehetmann. Multicenter evaluation of a fully automated high-throughput SARS-CoV-2 antigen immunoassay. . 2021; ():1.
Chicago/Turabian StyleDominik Nörz; Flaminia Olearo; Stojan Perisic; Matthias F. Bauer; Elena Riester; Tanja Schneider; Kathrin Schönfeld; Tina Laengin; Marc Lütgehetmann. 2021. "Multicenter evaluation of a fully automated high-throughput SARS-CoV-2 antigen immunoassay." , no. : 1.
So far, only a few reports about reinfections with SARS-CoV-2 have been published, and they often lack detailed immunological and virological data. We report about a SARS-CoV-2 reinfection with a genetically distinct SARS-CoV-2 variant in an immunocompetent female healthcare worker that has led to a mild disease course. No obvious viral escape mutations were observed in the second virus variant. The infectious virus was shed from the patient during the second infection episode despite the presence of neutralizing antibodies in her blood. Our data indicate that a moderate immune response after the first infection, but not a viral escape, did allow for reinfection and live virus shedding.
Thomas Brehm; Susanne Pfefferle; Ronald von Possel; Robin Kobbe; Dominik Nörz; Stefan Schmiedel; Adam Grundhoff; Flaminia Olearo; Petra Emmerich; Alexis Robitaille; Thomas Günther; Platon Braun; Gabriele Andersen; Johannes Knobloch; Marylyn Addo; Ansgar Lohse; Martin Aepfelbacher; Nicole Fischer; Julian Schulze Zur Wiesch; Marc Lütgehetmann. SARS-CoV-2 Reinfection in a Healthcare Worker Despite the Presence of Detectable Neutralizing Antibodies. Viruses 2021, 13, 661 .
AMA StyleThomas Brehm, Susanne Pfefferle, Ronald von Possel, Robin Kobbe, Dominik Nörz, Stefan Schmiedel, Adam Grundhoff, Flaminia Olearo, Petra Emmerich, Alexis Robitaille, Thomas Günther, Platon Braun, Gabriele Andersen, Johannes Knobloch, Marylyn Addo, Ansgar Lohse, Martin Aepfelbacher, Nicole Fischer, Julian Schulze Zur Wiesch, Marc Lütgehetmann. SARS-CoV-2 Reinfection in a Healthcare Worker Despite the Presence of Detectable Neutralizing Antibodies. Viruses. 2021; 13 (4):661.
Chicago/Turabian StyleThomas Brehm; Susanne Pfefferle; Ronald von Possel; Robin Kobbe; Dominik Nörz; Stefan Schmiedel; Adam Grundhoff; Flaminia Olearo; Petra Emmerich; Alexis Robitaille; Thomas Günther; Platon Braun; Gabriele Andersen; Johannes Knobloch; Marylyn Addo; Ansgar Lohse; Martin Aepfelbacher; Nicole Fischer; Julian Schulze Zur Wiesch; Marc Lütgehetmann. 2021. "SARS-CoV-2 Reinfection in a Healthcare Worker Despite the Presence of Detectable Neutralizing Antibodies." Viruses 13, no. 4: 661.
With great interest, we read the recent article by Zhen and Berry1Zhen W. Berry G.J. Development of a new multiplex real-time RT-PCR assay for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) detection.J Mol Diagn. 2020; 22: 1367-1372Abstract Full Text Full Text PDF PubMed Scopus (5) Google Scholar presenting a novel first-line diagnostic quantitative PCR assay for SARS-CoV-2 detection, targeting the viral S gene. Following reports of increasing abundance of SARS-CoV-2 variants, such as VOC202012/01 in Britain and 501.V2 in South Africa, a number of single nucleotide polymorphisms and in-frame deletions within the S gene have come into the spotlight in recent weeks.2Kemp S. Harvey W. Lytras S. Carabelli A. Robertson D. Gupta R. Recurrent emergence and transmission of a SARS-CoV-2 spike deletion H69/V70. bioRxiv. 2021..https://www.biorxiv.org/content/10.1101/2021.01.27.428516v1.full.pdfDate: 2021Google Scholar These mutations are increasingly attracting attention due to their potential consequences for infection control and vaccination programs.
Dominik Nörz; Susanne Pfefferle; Moritz Grunwald; Nicole Fischer; Martin Aepfelbacher; Marc Lütgehetmann. Modifying a Diagnostic SARS-CoV-2 Spike PCR to Turn a Del69/70 Dropout into a Discriminatory On-Target Assay. The Journal of Molecular Diagnostics 2021, 23, 777 -778.
AMA StyleDominik Nörz, Susanne Pfefferle, Moritz Grunwald, Nicole Fischer, Martin Aepfelbacher, Marc Lütgehetmann. Modifying a Diagnostic SARS-CoV-2 Spike PCR to Turn a Del69/70 Dropout into a Discriminatory On-Target Assay. The Journal of Molecular Diagnostics. 2021; 23 (6):777-778.
Chicago/Turabian StyleDominik Nörz; Susanne Pfefferle; Moritz Grunwald; Nicole Fischer; Martin Aepfelbacher; Marc Lütgehetmann. 2021. "Modifying a Diagnostic SARS-CoV-2 Spike PCR to Turn a Del69/70 Dropout into a Discriminatory On-Target Assay." The Journal of Molecular Diagnostics 23, no. 6: 777-778.
SARS-CoV-2 molecular diagnostics is facing material shortages and long turnaround times due to exponential increase of testing demand. We evaluated the analytic performance and handling of four rapid Antigen Point of Care Tests (AgPOCTs) I-IV (Distributors: (I) Roche, (II) Abbott, (III) MEDsan and (IV) Siemens). 100 RT-PCR negative and 84 RT-PCR positive oropharyngeal swabs were prospectively collected and used to determine performance and accuracy of these AgPOCTs. Handling was evaluated by 10 healthcare workers/users through a questionnaire. The median duration from symptom onset to sampling was 6 days (IQR 2–12 days). The overall respective sensitivity were 49.4 % (CI95 %: 38.9–59.9), 44.6 % (CI95 %: 34.3–55.3), 45.8 % (CI95 %: 35.5–56.5) and 54.9 % (CI95 %: 43.4−65.9) for tests I, II, III and IV, respectively. In the high viral load subgroup (containing >106 copies of SARS-CoV-2 /swab, n = 26), AgPOCTs reached sensitivities of 92.3 % or more (range 92.3 %–100 %). Specificity was 100 % for tests I, II (CI95 %: 96.3–100 for both tests) and IV (CI95 %: 96.3–100) and 97 % (CI95 %: 91.5–98.9) for test III. Regarding handling, test I obtained the overall highest scores, while test II was considered to have the most convenient components. Of note, users considered all assays, with the exception of test I, to pose a significant risk for contamination by drips or spills. Besides some differences in sensitivity and handling, all four AgPOCTs showed acceptable performance in high viral load samples. However, due to the significantly lower sensitivity compared to RT-qPCR, a careful consideration of pro and cons of AgPOCT has to be taken into account before clinical implementation.
Flaminia Olearo; Dominik Nörz; Fabian Heinrich; Jan Peter Sutter; Kevin Roedl; Alexander Schultze; Julian Schulze Zur Wiesch; Platon Braun; Lisa Oestereich; Benno Kreuels; Dominic Wichmann; Martin Aepfelbacher; Susanne Pfefferle; Marc Lütgehetmann. Handling and accuracy of four rapid antigen tests for the diagnosis of SARS-CoV-2 compared to RT-qPCR. Journal of Clinical Virology 2021, 137, 104782 -104782.
AMA StyleFlaminia Olearo, Dominik Nörz, Fabian Heinrich, Jan Peter Sutter, Kevin Roedl, Alexander Schultze, Julian Schulze Zur Wiesch, Platon Braun, Lisa Oestereich, Benno Kreuels, Dominic Wichmann, Martin Aepfelbacher, Susanne Pfefferle, Marc Lütgehetmann. Handling and accuracy of four rapid antigen tests for the diagnosis of SARS-CoV-2 compared to RT-qPCR. Journal of Clinical Virology. 2021; 137 ():104782-104782.
Chicago/Turabian StyleFlaminia Olearo; Dominik Nörz; Fabian Heinrich; Jan Peter Sutter; Kevin Roedl; Alexander Schultze; Julian Schulze Zur Wiesch; Platon Braun; Lisa Oestereich; Benno Kreuels; Dominic Wichmann; Martin Aepfelbacher; Susanne Pfefferle; Marc Lütgehetmann. 2021. "Handling and accuracy of four rapid antigen tests for the diagnosis of SARS-CoV-2 compared to RT-qPCR." Journal of Clinical Virology 137, no. : 104782-104782.
Hyperinflammation contributes to lung injury and subsequent acute respiratory distress syndrome with high mortality in patients with severe coronavirus disease 2019 (COVID-19). To understand the underlying mechanisms involved in lung pathology, we investigated the role of the lung-specific immune response. We profiled immune cells in bronchoalveolar lavage fluid and blood collected from patients with severe COVID-19 and patients with bacterial pneumonia not associated with viral infection. By tracking T cell clones across tissues, we identified clonally expanded tissue-resident memory-like TH17 cells (TRM17 cells) in the lungs even after viral clearance. These TRM17 cells were characterized by a potentially pathogenic cytokine expression profile of IL17A and CSF2 (GM-CSF). Interactome analysis suggests that TRM17 cells can interact with lung macrophages and cytotoxic CD8+ T cells, which have been associated with disease severity and lung damage. High IL-17A and GM-CSF protein levels in the serum of patients with COVID-19 were associated with a more severe clinical course. Collectively, our study suggests that pulmonary TRM17 cells are one potential orchestrator of the hyperinflammation in severe COVID-19.
Yu Zhao; Christoph Kilian; Jan-Eric Turner; Lidia Bosurgi; Kevin Roedl; Patricia Bartsch; Ann-Christin Gnirck; Filippo Cortesi; Christoph Schultheiß; Malte Hellmig; Leon U.B. Enk; Fabian Hausmann; Alina Borchers; Milagros N. Wong; Hans-Joachim Paust; Francesco Siracusa; Nicola Scheibel; Marissa Herrmann; Elisa Rosati; Petra Bacher; Dominik Kylies; Dominik Jarczak; Marc Lütgehetmann; Susanne Pfefferle; Stefan Steurer; Julian Schulze Zur Wiesch; Victor G. Puelles; Jan-Peter Sperhake; Marylyn M. Addo; Ansgar W. Lohse; Mascha Binder; Samuel Huber; Tobias B. Huber; Stefan Kluge; Stefan Bonn; Ulf Panzer; Nicola Gagliani; Christian F. Krebs. Clonal expansion and activation of tissue-resident memory-like TH17 cells expressing GM-CSF in the lungs of patients with severe COVID-19. Science Immunology 2021, 6, eabf6692 .
AMA StyleYu Zhao, Christoph Kilian, Jan-Eric Turner, Lidia Bosurgi, Kevin Roedl, Patricia Bartsch, Ann-Christin Gnirck, Filippo Cortesi, Christoph Schultheiß, Malte Hellmig, Leon U.B. Enk, Fabian Hausmann, Alina Borchers, Milagros N. Wong, Hans-Joachim Paust, Francesco Siracusa, Nicola Scheibel, Marissa Herrmann, Elisa Rosati, Petra Bacher, Dominik Kylies, Dominik Jarczak, Marc Lütgehetmann, Susanne Pfefferle, Stefan Steurer, Julian Schulze Zur Wiesch, Victor G. Puelles, Jan-Peter Sperhake, Marylyn M. Addo, Ansgar W. Lohse, Mascha Binder, Samuel Huber, Tobias B. Huber, Stefan Kluge, Stefan Bonn, Ulf Panzer, Nicola Gagliani, Christian F. Krebs. Clonal expansion and activation of tissue-resident memory-like TH17 cells expressing GM-CSF in the lungs of patients with severe COVID-19. Science Immunology. 2021; 6 (56):eabf6692.
Chicago/Turabian StyleYu Zhao; Christoph Kilian; Jan-Eric Turner; Lidia Bosurgi; Kevin Roedl; Patricia Bartsch; Ann-Christin Gnirck; Filippo Cortesi; Christoph Schultheiß; Malte Hellmig; Leon U.B. Enk; Fabian Hausmann; Alina Borchers; Milagros N. Wong; Hans-Joachim Paust; Francesco Siracusa; Nicola Scheibel; Marissa Herrmann; Elisa Rosati; Petra Bacher; Dominik Kylies; Dominik Jarczak; Marc Lütgehetmann; Susanne Pfefferle; Stefan Steurer; Julian Schulze Zur Wiesch; Victor G. Puelles; Jan-Peter Sperhake; Marylyn M. Addo; Ansgar W. Lohse; Mascha Binder; Samuel Huber; Tobias B. Huber; Stefan Kluge; Stefan Bonn; Ulf Panzer; Nicola Gagliani; Christian F. Krebs. 2021. "Clonal expansion and activation of tissue-resident memory-like TH17 cells expressing GM-CSF in the lungs of patients with severe COVID-19." Science Immunology 6, no. 56: eabf6692.
1 Abstract Background New SARS-CoV-2 variants with increased transmissibility, like B.1.1.7 from England or B1.351 from South Africa, have caused considerable concern worldwide. In order to contain the spread of these lineages, it is of utmost importance to have rapid, sensitive and high-throughput detection methods at hand. Methods Analytical sensitivity was assessed for both wild-type SARS-CoV-2 and B.1.1.7 lineage by serial dilution. A total of 141 clinical samples were subjected to the test and results compared to a commercial manual typing-PCR assay and NGS. Results The multiplex assay is highly sensitive for detection of SARS-CoV-2 RNA in clinical samples, with an LoD of 25.82 cp/ml (CI: 11.61 – 57.48). LoDs are slightly higher for the HV68/70 deletion (111.36 cp/ml; CI: 78.16 – 158.67) and the N501Y SNP (2548.04 cp/ml, CI: 1592.58 – 4076.73). A total of 141 clinical samples were tested with the assay, including 16 samples containing SARS-CoV-2 of the B.1.1.7 lineage. Three non-B.1.1.7 samples contained a HV69/70 deletion. All were correctly identified by the multiplex assay. Conclusion We describe here a highly sensitive, fully automated multiplex PCR assay for the simultaneous detection of del-HV69/70 and N501Y that can distinguish between lineages B.1.1.7 and B1.351. The assay allows for high-throughput screening for relevant variants in clinical samples prior to sequencing.
Dominik Nörz; Moritz Grunwald; Flaminia Olearo; Nicole Fischer; Martin Aepfelbacher; Susanne Pfefferle; Marc Lütgehetmann. Evaluation of a fully automated high-throughput SARS-CoV-2 multiplex qPCR assay with build-in screening functionality for DelHV69/70- and N501Y variants such as B.1.1.7. 2021, 1 .
AMA StyleDominik Nörz, Moritz Grunwald, Flaminia Olearo, Nicole Fischer, Martin Aepfelbacher, Susanne Pfefferle, Marc Lütgehetmann. Evaluation of a fully automated high-throughput SARS-CoV-2 multiplex qPCR assay with build-in screening functionality for DelHV69/70- and N501Y variants such as B.1.1.7. . 2021; ():1.
Chicago/Turabian StyleDominik Nörz; Moritz Grunwald; Flaminia Olearo; Nicole Fischer; Martin Aepfelbacher; Susanne Pfefferle; Marc Lütgehetmann. 2021. "Evaluation of a fully automated high-throughput SARS-CoV-2 multiplex qPCR assay with build-in screening functionality for DelHV69/70- and N501Y variants such as B.1.1.7." , no. : 1.
In patients with hepatitis E virus (HEV) infections, extrahepatic, particularly renal and hematological manifestations, are increasingly reported in the medical literature but have never been studied compared to a control cohort. We retrospectively analyzed medical records of consecutive patients that were diagnosed with acute hepatitis E (AHE) (n = 69) or acute hepatitis A (AHA) (n = 46) at the University Medical Center Hamburg Eppendorf from January 2009 to August 2019 for demographical, clinical, and laboratory information. Patients with AHE had significantly lower median levels of ALAT (798 U/L) and total bilirubin (1.8 mg/dL) compared to patients with AHA (2326 U/L; p < 0.001 and 5.2 mg/dL; p < 0.001), suggesting a generally less severe hepatitis. In contrast, patients with AHE had significantly higher median serum creatinine levels (0.9 mg/dL vs. 0.8 mg/dL; p = 0.002) and lower median estimated glomerular filtration rate (eGFR) (91 mL/min/1.73 m2 vs. 109 mL/min/1.73 m2; p < 0.001) than patients with AHA. Leucocyte, neutrophil and lymphocyte count, hemoglobin, platelets, red cell distribution width (RDW), neutrophil to lymphocyte ratio (NLR), and RDW to lymphocyte ratio (RLR) did not differ between patients with AHE and those with AHA. Our observations indicate that renal but not hematological interference presents an underrecognized extrahepatic feature of AHE, while inflammation of the liver seems to be more severe in AHA.
Thomas Theo Brehm; Omid Mazaheri; Thomas Horvatits; Marc Lütgehetmann; Julian Schulze Zur Wiesch; Ansgar W. Lohse; Susanne Polywka; Sven Pischke. Lower Levels of Transaminases but Higher Levels of Serum Creatinine in Patients with Acute Hepatitis E in Comparison to Patients with Hepatitis A. Pathogens 2021, 10, 60 .
AMA StyleThomas Theo Brehm, Omid Mazaheri, Thomas Horvatits, Marc Lütgehetmann, Julian Schulze Zur Wiesch, Ansgar W. Lohse, Susanne Polywka, Sven Pischke. Lower Levels of Transaminases but Higher Levels of Serum Creatinine in Patients with Acute Hepatitis E in Comparison to Patients with Hepatitis A. Pathogens. 2021; 10 (1):60.
Chicago/Turabian StyleThomas Theo Brehm; Omid Mazaheri; Thomas Horvatits; Marc Lütgehetmann; Julian Schulze Zur Wiesch; Ansgar W. Lohse; Susanne Polywka; Sven Pischke. 2021. "Lower Levels of Transaminases but Higher Levels of Serum Creatinine in Patients with Acute Hepatitis E in Comparison to Patients with Hepatitis A." Pathogens 10, no. 1: 60.
Background: SARS-CoV-2 molecular diagnostics is facing material shortages and long turnaround times due to exponential increase of testing demand. Objective: We evaluated the analytic performance and handling of four rapid Antigen Point of Care Tests (AgPOCTs) I-IV (Distributors: (I) Roche, (II) Abbott, (III) MEDsan and (IV) Siemens). Methods: 100 RT-PCR negative and 84 RT-PCR positive oropharyngeal swabs were prospectively collected and used to determine performance and accuracy of these AgPOCTs. Handling was evaluated by 10 healthcare workers/users through a questionnaire. Results: The median duration from symptom onset to sampling was 6 days (IQR 2-12 days). The overall relative sensitivity was 49.4%, 44.6%, 45.8% and 54.9 % for tests I, II, III and IV, respectively. In the high viral load subgroup (containing >106 copies of SARS-CoV-2 /swab, n=26), AgPOCTs reached sensitivities of 92.3% or more (range 92.3%-100%). Specificity was 100% for tests I, II and IV and 97% for test III. Regarding handling, test I obtained the overall highest scores, while test II was considered to have the most convenient components. Of note, users considered all assays, with the exception of test I, to pose a significant risk for contamination by drips or spills. Discussion: Besides some differences in sensitivity and handling, all four AgPOCTs showed acceptable performance in high viral load samples. However, due to the significantly lower sensitivity compared to RT-qPCR, a careful consideration of pro and cons of AgPOCT has to be taken into account before clinical implementation.
Flaminia Olearo; Dominik Noerz; Fabian Heinrich; Jan Peter Sutter; Kevin Roedel; Alexander Schultze; Julian Schulze Zur Wiesch; Platon Braun; Lisa Oesterreich; Benno Kreuels; Dominic Wichmann; Martin Aepfelbacher; Susanne Pfefferle; Marc Luetgehetmann. Handling and accuracy of four rapid antigen tests for the diagnosis of SARS-CoV-2 compared to RT-qPCR. 2020, 1 .
AMA StyleFlaminia Olearo, Dominik Noerz, Fabian Heinrich, Jan Peter Sutter, Kevin Roedel, Alexander Schultze, Julian Schulze Zur Wiesch, Platon Braun, Lisa Oesterreich, Benno Kreuels, Dominic Wichmann, Martin Aepfelbacher, Susanne Pfefferle, Marc Luetgehetmann. Handling and accuracy of four rapid antigen tests for the diagnosis of SARS-CoV-2 compared to RT-qPCR. . 2020; ():1.
Chicago/Turabian StyleFlaminia Olearo; Dominik Noerz; Fabian Heinrich; Jan Peter Sutter; Kevin Roedel; Alexander Schultze; Julian Schulze Zur Wiesch; Platon Braun; Lisa Oesterreich; Benno Kreuels; Dominic Wichmann; Martin Aepfelbacher; Susanne Pfefferle; Marc Luetgehetmann. 2020. "Handling and accuracy of four rapid antigen tests for the diagnosis of SARS-CoV-2 compared to RT-qPCR." , no. : 1.
Despite improvements in diagnosis, intensive-care medicine and surgical technique, the mortality of patients with secondary peritonitis is still high. Early and aggressive empiric antibiotic treatment has strong impact on the outcome. This retrospective study investigates bacterial and fungal pathogens and their antibiotic sensitivity in patients with secondary peritonitis. All patients that underwent emergency laparotomy due to secondary peritonitis at the Department of Surgery, University Medical Center Hamburg-Eppendorf between 2005 and 2015 were reviewed and overall 414 patients were included. We correlated the intra-abdominal localization of the organ perforation with intraoperative microbiological findings and corresponding sensitivities to relevant antibiotics. Overall, the most common findings were Escherichia coli (39%) and other Enterobacterica (24%). Depending on the location of the perforation, Cefuroxime/Metronidazole and Cefutaxime/Metronidazole were effective (based on in vitro susceptibility testing) in only 55–73% of the patients, while Meropenem/Vancomycin was able to control the peritonitis in more than 98% of the patients; independent of the location. Besides early source control, appropriate empiric treatment plays a pivotal role in treatment of secondary peritonitis. We are able to show that the frequently used combinations of second or third generation Cephalosporins with Metronidazole are not always sufficient, which is due to the biological resistance of the bacteria. Further clinical studies are needed to determine whether calculated use of broad-spectrum antibiotics with a sensitivity rate > 99%, such as Carbapenem plus Vancomycin, can improve overall survival rates in critically ill patients with secondary peritonitis.
Rainer Grotelüschen; Lena M. Heidelmann; Marc Lütgehetmann; Nathaniel Melling; Matthias Reeh; Tarik Ghadban; Anna Dupree; Jakob R. Izbicki; Kai A. Bachmann. Antibiotic sensitivity in correlation to the origin of secondary peritonitis: a single center analysis. Scientific Reports 2020, 10, 1 -9.
AMA StyleRainer Grotelüschen, Lena M. Heidelmann, Marc Lütgehetmann, Nathaniel Melling, Matthias Reeh, Tarik Ghadban, Anna Dupree, Jakob R. Izbicki, Kai A. Bachmann. Antibiotic sensitivity in correlation to the origin of secondary peritonitis: a single center analysis. Scientific Reports. 2020; 10 (1):1-9.
Chicago/Turabian StyleRainer Grotelüschen; Lena M. Heidelmann; Marc Lütgehetmann; Nathaniel Melling; Matthias Reeh; Tarik Ghadban; Anna Dupree; Jakob R. Izbicki; Kai A. Bachmann. 2020. "Antibiotic sensitivity in correlation to the origin of secondary peritonitis: a single center analysis." Scientific Reports 10, no. 1: 1-9.
Objectives Investigation whether in depth characterization of virus variant patterns can be used for epidemiological analysis of the first severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection clusters in Hamburg, Germany. Methods Metagenomic RNA-sequencing and amplicon-sequencing and subsequent variant calling in 25 respiratory samples from SARS-CoV-2 infected patients involved in the earliest infection clusters in Hamburg. Results Amplikon sequencing and cluster analyses of these SARS-CoV-2 sequences allowed the identification of the first infection cluster and five non-related infection clusters occurring at the beginning of the viral entry of SARS-CoV-2 in the Hamburg metropolitan region. Viral genomics together with epidemiological analyses revealed that the index patient acquired the infection in northern Italy and transmitted it to two out of 134 contacts. Single nucleotide polymorphisms clearly distinguished the virus variants of the index and other clusters and allowed us to track in which sequences worldwide these mutations were first described. Minor variant analyses identified the transmission of intra-host variants in the index cluster and household clusters. Conclusions SARS-CoV-2 variant tracing allows the identification of infection clusters and the follow up of infection chains occurring in the population. Furthermore, the follow up of minor viral variants in infection clusters can provide further resolution on transmission events indistinguishable at a consensus sequence level.
Susanne Pfefferle; Thomas Günther; Robin Kobbe; Manja Czech-Sioli; Dominic Nörz; René Santer; Jun Oh; Stefan Kluge; Lisa Oestereich; Kersten Peldschus; Daniela Indenbirken; Jiabin Huang; Adam Grundhoff; Martin Aepfelbacher; Johannes K. Knobloch; Marc Lütgehetmann; Nicole Fischer. SARS Coronavirus-2 variant tracing within the first Coronavirus Disease 19 clusters in northern Germany. Clinical Microbiology and Infection 2020, 27, 130.e5 -130.e8.
AMA StyleSusanne Pfefferle, Thomas Günther, Robin Kobbe, Manja Czech-Sioli, Dominic Nörz, René Santer, Jun Oh, Stefan Kluge, Lisa Oestereich, Kersten Peldschus, Daniela Indenbirken, Jiabin Huang, Adam Grundhoff, Martin Aepfelbacher, Johannes K. Knobloch, Marc Lütgehetmann, Nicole Fischer. SARS Coronavirus-2 variant tracing within the first Coronavirus Disease 19 clusters in northern Germany. Clinical Microbiology and Infection. 2020; 27 (1):130.e5-130.e8.
Chicago/Turabian StyleSusanne Pfefferle; Thomas Günther; Robin Kobbe; Manja Czech-Sioli; Dominic Nörz; René Santer; Jun Oh; Stefan Kluge; Lisa Oestereich; Kersten Peldschus; Daniela Indenbirken; Jiabin Huang; Adam Grundhoff; Martin Aepfelbacher; Johannes K. Knobloch; Marc Lütgehetmann; Nicole Fischer. 2020. "SARS Coronavirus-2 variant tracing within the first Coronavirus Disease 19 clusters in northern Germany." Clinical Microbiology and Infection 27, no. 1: 130.e5-130.e8.
The ongoing SARS-CoV-2 pandemic presents a unique challenge to diagnostic laboratories. There are preliminary studies correlating qRT-PCR results from different materials to clinical outcomes, yet, comparability is limited due to the plethora of different assays used for diagnostics. In this study we evaluate clinical performance and linear range for the SARS-CoV-2 IVD (cobas6800/8800 system, a fully automated sample-to-result platform) in different clinically relevant matrix materials outside official specifications. Assay performance was assessed in human plasma, BAL/BL and transport medium following chemical inactivation. For analytical evaluation, respective matrix materials were spiked with SARS-CoV-2 RNA in ten-fold dilution series. The efficacy of chemical inactivation by guanidine hydrochloride solution was confirmed in cell culture infectivity experiments. For correlation, a total of 289 predetermined clinical samples including respiratory swabs, plasma and lower respiratory tract specimens were subjected to the SARS-CoV-2 IVD test and results were compared. The SARS-CoV-2 IVD showed excellent linearity over four to six log steps depending on matrix material. Chemical inactivation resulted in a reduction in plaque forming units of at least 3.5 log steps, while having no significant impact on assay performance. Inter-run consistency from three different testing sites demonstrated excellent comparability of RT-PCR results (maximum deviation was 1.53 CT). Clinical evaluation for respiratory swabs showed very good agreement with the comparator assay (Positive agreement 95.7 %, negative agreement 98.9 %). The SARS-CoV-2 IVD test for the cobas6800/8800 systems offers excellent linear range and inter-run consistency for quantification of SARS-CoV-2 RNA in different matrices outside official specifications.
Dominik Nörz; André Frontzek; Ulrich Eigner; Lisa Oestereich; Dominic Wichmann; Stefan Kluge; Nicole Fischer; Martin Aepfelbacher; Susanne Pfefferle; Marc Lütgehetmann. Pushing beyond specifications: Evaluation of linearity and clinical performance of the cobas 6800/8800 SARS-CoV-2 RT-PCR assay for reliable quantification in blood and other materials outside recommendations. Journal of Clinical Virology 2020, 132, 104650 -104650.
AMA StyleDominik Nörz, André Frontzek, Ulrich Eigner, Lisa Oestereich, Dominic Wichmann, Stefan Kluge, Nicole Fischer, Martin Aepfelbacher, Susanne Pfefferle, Marc Lütgehetmann. Pushing beyond specifications: Evaluation of linearity and clinical performance of the cobas 6800/8800 SARS-CoV-2 RT-PCR assay for reliable quantification in blood and other materials outside recommendations. Journal of Clinical Virology. 2020; 132 ():104650-104650.
Chicago/Turabian StyleDominik Nörz; André Frontzek; Ulrich Eigner; Lisa Oestereich; Dominic Wichmann; Stefan Kluge; Nicole Fischer; Martin Aepfelbacher; Susanne Pfefferle; Marc Lütgehetmann. 2020. "Pushing beyond specifications: Evaluation of linearity and clinical performance of the cobas 6800/8800 SARS-CoV-2 RT-PCR assay for reliable quantification in blood and other materials outside recommendations." Journal of Clinical Virology 132, no. : 104650-104650.
Background: Hepatitis E virus (HEV) has been associated with immunological phenomena. Their clinical significance, however, still needs to be clarified, that is, whether cryoglobulins or autoantibodies impact overt disease in HEV-infected individuals. To better understand, we analyzed these different immune phenomena in three cohorts, each representing different types of HEV infection. Methods: The cohorts included: (i) immunocompetent patients with acute hepatitis E, (ii) immunosuppressed patients with chronic hepatitis E, and (iii) individuals with asymptomatic HEV infection. Together, they consisted of 57 individuals and were studied retrospectively for the presence of anti-nuclear antibodies (ANAs), cryoglobulins, and serum total IgG. They were then compared with a control cohort of 17 untreated patients with chronic hepatitis B virus (HBV) infection or hepatitis C virus (HCV) infection. Results: Thirteen (23%) were immunocompetent patients with acute hepatitis E (median alanine aminotransferase (ALT) = 872 U/L), 15 (26%) were immunosuppressed patients with chronic hepatitis E (median ALT = 137 U/L), and 29 (51%) were blood donors with asymptomatic HEV infection (median ALT = 35 U/L). Overall, 24% tested positive for elevated ANA titers of >1:160, and 11% presented with a specific ANA pattern. ANA detection was not associated with the type of HEV infection, IgG levels, sex, or age. All individuals tested negative for anti-mitochondrial antibodies, anti-neutrophil cytoplasmic antibodies, liver-kidney microsomal antibodies, anti-myeloperoxidase-, and anti-proteinase-3 antibodies. Five patients (9%) tested positive for cryoglobulins. Notably, cryoglobulinemia was present in overt hepatitis E (Groups (i) and (ii); one acute and four chronic HEV infections), but was not present in any of the asymptomatic blood donors (p = 0.02). The frequency of cryoglobulins and elevated ANAs did not differ significantly between HEV and HBV/HCV patients. Conclusion: In line with findings on HBV and HCV infections, we frequently observed detection of ANAs (24%) and cryoglobulins (9%) in association with HEV infections. The presence of cryoglobulins was limited to patients with overt hepatitis E. We add to the findings on the immune phenomena of hepatitis E.
Thomas Horvatits; Julian Schulze Zur Wiesch; Susanne Polywka; Gustav Buescher; Marc Lütgehetmann; Elaine Hussey; Karoline Horvatits; Sven Peine; Friedrich Haag; Marylyn M. Addo; Ansgar W. Lohse; Christina Weiler-Normann; Sven Pischke. Significance of Anti-Nuclear Antibodies and Cryoglobulins in Patients with Acute and Chronic HEV Infection. Pathogens 2020, 9, 755 .
AMA StyleThomas Horvatits, Julian Schulze Zur Wiesch, Susanne Polywka, Gustav Buescher, Marc Lütgehetmann, Elaine Hussey, Karoline Horvatits, Sven Peine, Friedrich Haag, Marylyn M. Addo, Ansgar W. Lohse, Christina Weiler-Normann, Sven Pischke. Significance of Anti-Nuclear Antibodies and Cryoglobulins in Patients with Acute and Chronic HEV Infection. Pathogens. 2020; 9 (9):755.
Chicago/Turabian StyleThomas Horvatits; Julian Schulze Zur Wiesch; Susanne Polywka; Gustav Buescher; Marc Lütgehetmann; Elaine Hussey; Karoline Horvatits; Sven Peine; Friedrich Haag; Marylyn M. Addo; Ansgar W. Lohse; Christina Weiler-Normann; Sven Pischke. 2020. "Significance of Anti-Nuclear Antibodies and Cryoglobulins in Patients with Acute and Chronic HEV Infection." Pathogens 9, no. 9: 755.
In vitro cell culture experiments and animal models have demonstrated that hepatitis delta virus (HDV) can theoretically propagate being enveloped by human pathogenic viruses other than hepatitis B virus (HBV), namely hepatitis C virus (HCV) and dengue virus. However, the clinical relevance of these findings, and whether HDV replication occurs in real‐world hepatitis B surface antigen (HBsAg) negative HCV patient cohorts remains unknown. To this aim, we analysed 323 HCV‐RNA positive and HBsAg negative sera for the presence of HDV‐RNA and anti‐HDV antibodies (anti‐HDV). All 323 (100%) samples were negative for HDV‐RNA. Interestingly, 8/316 samples tested positive for anti‐HDV. The HBV‐serology of these eight patients showed a positive result for HBV core antibodies (anti‐HBc) indicating a seroconversion of an acute HBV infection in the past. None of the anti‐HBc negative patients were positive for anti‐HDV. Our results indicate a distinctly low probability of replicative HDV infection in HCV mono‐infected patients in Germany. Current German clinical guidelines rightly recommend performing HDV screening only in HBsAg positive patients. However, larger studies on this subject should be performed in regions that are endemic for chronic HBV/HDV as well as HCV infections.
Lisa Sophie Pflüger; Julian Schulze Zur Wiesch; Susanne Polywka; Marc Lütgehetmann. Hepatitis delta virus propagation enabled by hepatitis C virus—Scientifically intriguing, but is it relevant to clinical practice? Journal of Viral Hepatitis 2020, 28, 213 -216.
AMA StyleLisa Sophie Pflüger, Julian Schulze Zur Wiesch, Susanne Polywka, Marc Lütgehetmann. Hepatitis delta virus propagation enabled by hepatitis C virus—Scientifically intriguing, but is it relevant to clinical practice? Journal of Viral Hepatitis. 2020; 28 (1):213-216.
Chicago/Turabian StyleLisa Sophie Pflüger; Julian Schulze Zur Wiesch; Susanne Polywka; Marc Lütgehetmann. 2020. "Hepatitis delta virus propagation enabled by hepatitis C virus—Scientifically intriguing, but is it relevant to clinical practice?" Journal of Viral Hepatitis 28, no. 1: 213-216.