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Fast SARS-CoV-2 detection by RT-qPCR in preheated nasopharyngeal swab samples

OBJECTIVES: The gold-standard COVID-19 diagnosis relies on detecting SARS-CoV-2 using RNA purification and one-step retrotranscription and quantitative PCR (RT-qPCR). Based on the urgent need for high-throughput screening, we tested the performance of three alternative, simple and affordable protocols to rapidly detect SARS-CoV-2, bypassing the long and tedious RNA extraction step and reducing the time to viral detection

METHODS: We evaluated three methods based on direct nasopharyngeal swab viral transmission medium (VTM) heating before the RT-qPCR: a) direct without additives; b) in a formamide-EDTA (FAE) buffer, c) in a RNAsnapTM buffer

RESULTS: Although with a delay in cycle threshold compared to the gold-standard, we found consistent results in nasopharyngeal swab samples that were subject to a direct 70°C incubation for 10 min

CONCLUSIONS: Our findings provide valuable options to overcome any supply chain issue and help to increase the throughput of diagnostic tests, thereby complementing standard diagnosis

Year of Publication: 2020
Contained in: International journal of infectious diseases : IJID : official publication of the International Society for Infectious Diseases Vol. 97 (2020), p. 66-68
All journal articles: Search for all articles in this journal
Language: English
Contributors: Alcoba-Florez, Julia | Author
González-Montelongo, Rafaela
Íñigo-Campos, Antonio
de Artola, Diego García-Martínez
Gil-Campesino, Helena
The Microbiology Technical Support Team
Ciuffreda, Laura
Valenzuela-Fernández, Agustín
Flores, Carlos
Full text access:
Electronic availability is being checked...
Links: Full Text (dx.doi.org)
Keywords: COVID-19
Journal Article
RNA extraction
SARS-CoV-2
diagnosis
fast protocols
sample treatment
Additional Keywords: Betacoronavirus
Coronavirus Infections
Diagnostic Tests, Routine
Hot Temperature
Humans
Pandemics
Pneumonia, Viral
Real-Time Polymerase Chain Reaction
ISSN: 1878-3511
Note: Copyright: From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Notes: Date Completed 03.08.2020
Date Revised 03.08.2020
published: Print-Electronic
Citation Status MEDLINE
Copyright: From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
PMID:
    32492531
Physical Description: Online-Ressource
ID (e.g. DOI, URN): 10.1016/j.ijid.2020.05.099
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520 |a METHODS: We evaluated three methods based on direct nasopharyngeal swab viral transmission medium (VTM) heating before the RT-qPCR: a) direct without additives; b) in a formamide-EDTA (FAE) buffer, c) in a RNAsnapTM buffer 
520 |a RESULTS: Although with a delay in cycle threshold compared to the gold-standard, we found consistent results in nasopharyngeal swab samples that were subject to a direct 70°C incubation for 10 min 
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