Serology- and PCR-based cumulative incidence of SARS-CoV-2 infection in adults in a successfully contained early hotspot (CoMoLo study), Germany, May to June 2020

Santos-Hövener, Claudia; Neuhauser, Hannelore; Rosario, Angelika Schaffrath; Busch, Markus; Schlaud, Martin; Hoffmann, Robert; Gößwald, Antje; Koschollek, Carmen; Hoebel, Jens; Allen, Jennifer; Haack-Erdmann, Antje; Brockmann, Stefan; Ziese, Thomas; Nitsche, Andreas; Michel, Janine; Haller, Sebastian; Wilking, Hendrik; Hamouda, Osamah; Corman, Victor M.; Drosten, Christian; Schaade, Lars; Wieler, Lothar H.; Lampert, Thomas

doi:10.2807/1560-7917.es.2020.25.47.2001752

Cited by 70 publications

(73 citation statements)

References 24 publications

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“…At present, the United States Centers for Disease Control and Prevention (CDC) recommends diagnosis of acute SARS-CoV-2 infection via measurement of viral nucleic acid and antigen tests ( https://www.cdc.gov/coronavirus/2019-nCoV/hcp/clinical-criteria.html ). Real-time reverse transcriptase-polymerase chain reaction (real-time RT-PCR) is a reliable and relatively fast method for the identification of pathogenic nucleic acids in patient samples 6 . However, it has several disadvantages.…”

Section: Introductionmentioning

confidence: 99%

“…Another disadvantage is the requirement for relatively high concentrations of genetic material in a sample. Although real-time RT-PCR is considered to be a sensitive method for the detection of nucleic acid, the limit of detection for SARS-CoV-2 RNA is reported to be between 200 and 77,440 copies/mL, depending on the RNA extraction method and real-time RT-PCR assay being used 6 , 9 , 10 .…”

Section: Introductionmentioning

confidence: 99%

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Hydrogel particles improve detection of SARS-CoV-2 RNA from multiple sample types

Barclay

Akhrymuk

Patnaik

et al. 2020

Sci Rep

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Here we present a rapid and versatile method for capturing and concentrating SARS-CoV-2 from contrived transport medium and saliva samples using affinity-capture magnetic hydrogel particles. We demonstrate that the method concentrates virus from 1 mL samples prior to RNA extraction, substantially improving detection of virus using real-time RT-PCR across a range of viral titers (100–1,000,000 viral copies/mL) and enabling detection of virus using the 2019 nCoV CDC EUA Kit down to 100 viral copies/mL. This method is compatible with commercially available nucleic acid extraction kits (i.e., from Qiagen) and a simple heat and detergent method that extracts viral RNA directly off the particle, allowing a sample processing time of 10 min. We furthermore tested our method in transport medium diagnostic remnant samples that previously had been tested for SARS-CoV-2, showing that our method not only correctly identified all positive samples but also substantially improved detection of the virus in low viral load samples. The average improvement in cycle threshold value across all viral titers tested was 3.1. Finally, we illustrate that our method could potentially be used to enable pooled testing, as we observed considerable improvement in the detection of SARS-CoV-2 RNA from sample volumes of up to 10 mL.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Hydrogel particles improve detection of SARS-CoV-2 RNA from multiple sample types

Barclay

Akhrymuk

Patnaik

et al. 2020

Sci Rep

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“…Das E (Hüllprotein)-Gen ist ein stark konserviertes Gen, das in SARS und in SARS-CoV‑2 identisch ist (pan-Sarbecovirus-Gen). Daher wird in einem Protokoll empfohlen, die E (Hüllprotein)-Genamplifikation als Vortest für den Nachweis von SARS-Viren zu verwenden und im Falle eines positiven Ergebnisses anschließend 1 oder 2 weitere Gene von SARS-CoV‑2 als Bestätigungstest zu amplifizieren ( N -[Nukleokapsidprotein]-Gen, RdRp [RNA-abhängiges RNA-Polymerase]-Gen) [ 10 ]. Das S (Spikeprotein)-Gen weist aufgrund von hohem Selektionsdruck häufiger Mutationen auf, weshalb die alleinige Amplifikation des S -Gens nicht empfohlen wird.…”

Section: Sars-cov-2-nachweismethodsenunclassified

Nachweismethoden von SARS-CoV-2 in Gewebe

Stillfried

2021

Pathologe

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Hintergrund Die Analyse von SARS-CoV‑2 in Geweben von COVID-19-Patienten ist wichtig für ein besseres Verständnis der Pathophysiologie der Krankheit, die Interpretation der diagnostischen histopathologischen Befunde in Autopsien, Biopsien und Resektaten oder um ein potenzielles berufsbedingtes Infektionsrisiko einzuschätzen. Material und Methoden In dieser Übersichtsarbeit haben wir 136 publizierte Studien zu Detektionsmethoden von SARS-CoV‑2 in Gewebe in der kuratierten Literaturdatenbank LitCovid von PubMed identifiziert und bezüglich Fehlerquellen, Spezifität und Sensitivität der Methoden unter Berücksichtigung eigener Erfahrungen ausgewertet. Ergebnisse Es gibt keine ausreichend spezifischen histomorphologischen Veränderungen bzw. diagnostischen Merkmale von COVID-19. Daher werden 3 Ansätze zum SARS-CoV-2-Nachweis genutzt: Nachweis von RNA, Proteinen/Antigenen oder morphologischer Nachweis mittels Elektronenmikroskopie. In der präanalytischen Phase liegt die dominante Fehlerquelle in der Gewebequalität, insbesondere den unterschiedlichen Intervallen zwischen Probenentnahme und -aufarbeitung, sowie spezifisch in Autopsien im Intervall zwischen Tod und Probenentnahme. Diese Angaben finden sich in weniger als der Hälfte der Studien (z. B. nur in 42 % der Autopsiestudien). Eigene Erfahrungen und erste Studien belegen die deutlich höhere Sensitivität und Spezifität von RNA-basierten Nachweismethoden gegenüber Antigen- bzw. Proteinnachweis mittels Immunhistochemie oder Immunfluoreszenz. Der Nachweis mittels Elektronenmikroskopie ist zeitintensiv und die Interpretation schwierig. Schlussfolgerungen Es stehen verschiedene Methoden zum Nachweis von SARS-CoV‑2 im Gewebe zur Verfügung. Derzeit ist der RNA-Nachweis mittels RT-PCR die Methode der Wahl. Notwendige, umfangreiche Validationsstudien und Methodenharmonisierung sind derzeit noch nicht verfügbar. Zusatzmaterial online Zusätzliche Informationen sind in der Online-Version dieses Artikels (10.1007/s00292-021-00919-8) enthalten.

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“…Molecular techniques have been designed to address this need 9 . According to the World Health Organization (WHO), the gold standard method to detect SARS-CoV-2 is real-time polymerase chain reaction (RT-PCR) using TaqMan probes, which precisely detect the presence of the virus 10 . However, due to the intensive labor required to perform the technique, the reagents involved, and the limited availability of kits, many diagnoses are based only on late-stage symptoms 11 .…”

Section: Introductionmentioning

confidence: 99%

A new SYBR Green real-time PCR to detect SARS-CoV-2

Marinowic

Zanirati

Rodrigues

et al. 2021

Sci Rep

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Phylogenetic analysis has demonstrated that the etiologic agent of the 2020 pandemic outbreak is a betacoronavirus named SARS-CoV-2. For public health interventions, a diagnostic test with high sensitivity and specificity is required. The gold standard protocol for diagnosis by the Word Health Organization (WHO) is RT-PCR. To detect low viral loads and perform large-scale screening, a low-cost diagnostic test is necessary. Here, we developed a cost-effective test capable of detecting SARS-CoV-2. We validated an auxiliary protocol for molecular diagnosis with the SYBR Green RT-PCR methodology to successfully screen negative cases of SARS-CoV-2. Our results revealed a set of primers with high specificity and no homology with other viruses from the Coronovideae family or human respiratory tract pathogenic viruses, presenting with complementarity only for rhinoviruses/enteroviruses and Legionella spp. Optimization of the annealing temperature and polymerization time led to a high specificity in the PCR products. We have developed a more affordable and swift methodology for negative SARS-CoV-2 screening. This methodology can be applied on a large scale to soften panic and economic burden through guidance for isolation strategies.

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Serology- and PCR-based cumulative incidence of SARS-CoV-2 infection in adults in a successfully contained early hotspot (CoMoLo study), Germany, May to June 2020

Cited by 70 publications

References 24 publications

Hydrogel particles improve detection of SARS-CoV-2 RNA from multiple sample types

Hydrogel particles improve detection of SARS-CoV-2 RNA from multiple sample types

Nachweismethoden von SARS-CoV-2 in Gewebe

A new SYBR Green real-time PCR to detect SARS-CoV-2

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