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A Streamlined Approach to Rapidly Detect SARS-CoV-2 Infection Avoiding RNA Extraction: Workflow Validation
- Source :
- Disease Markers, Vol 2020 (2020), Disease Markers
- Publication Year :
- 2020
- Publisher :
- Hindawi Limited, 2020.
-
Abstract
- Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection has rapidly spread worldwide from the beginning of 2020. The presence of viral RNA in samples by nucleic acid (NA) molecular analysis is the only method available to diagnose COVID-19 disease and to assess patients’ viral load. Since the demand for laboratory reagents has increased, there has been a worldwide shortage of RNA extraction kits. We, therefore, developed a fast and cost-effective viral genome isolation method that, combined with quantitative RT-PCR assay, detects SARS-CoV-2 RNA in patient samples. The method relies on the addition of Proteinase K followed by a controlled heat-shock incubation and, then, E gene evaluation by RT-qPCR. It was validated for sensitivity, specificity, linearity, reproducibility, and precision. It detects as low as 10 viral copies/sample, is rapid, and has been characterized in 60 COVID-19-infected patients. Compared to automated extraction methods, our pretreatment guarantees the same positivity rate with the advantage of shortening the time of the analysis and reducing its cost. This is a rapid workflow meant to aid the healthcare system in the rapid identification of infected patients, such as during a pathogen-related outbreak. For its intrinsic characteristics, this workflow is suitable for large-scale screenings.
- Subjects :
- Medicine (General)
Article Subject
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)
Clinical Biochemistry
02 engineering and technology
Sensitivity and Specificity
Workflow
Coronavirus Envelope Proteins
03 medical and health sciences
COVID-19 Testing
R5-920
0302 clinical medicine
Limit of Detection
Nasopharynx
Genetics
Humans
Medicine
030212 general & internal medicine
Molecular Biology
Gene
biology
Reverse Transcriptase Polymerase Chain Reaction
SARS-CoV-2
business.industry
Biochemistry (medical)
RNA
General Medicine
021001 nanoscience & nanotechnology
Proteinase K
Virology
Nucleic acid
biology.protein
RNA, Viral
RNA extraction
0210 nano-technology
business
Viral load
Research Article
Subjects
Details
- ISSN :
- 18758630 and 02780240
- Volume :
- 2020
- Database :
- OpenAIRE
- Journal :
- Disease Markers
- Accession number :
- edsair.doi.dedup.....491d94b674f43a17a20db94588873c31