Portable and accurate diagnostics for COVID-19: Combined use of the miniPCR thermocycler and a well-plate reader for SARS-CoV-2 virus detection

The coronavirus disease 2019 (COVID-19) pandemic has crudely demonstrated the need for massive and rapid diagnostics. By the first week of July, more than 10,000,000 positive cases of COVID-19 have been reported worldwide, although this number could be greatly underestimated. In the case of an epide...

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Bibliographic Details
Published in:PloS one 2020-08, Vol.15 (8), p.e0237418-e0237418
Main Authors: González-González, Everardo, Trujillo-de Santiago, Grissel, Lara-Mayorga, Itzel Montserrat, Martínez-Chapa, Sergio Omar, Alvarez, Mario Moisés
Format: Article
Language:English
Subjects:
Amplification
Base Sequence
Betacoronavirus - chemistry
Betacoronavirus - genetics
Biology and Life Sciences
Coronaviridae
Coronavirus Infections - diagnosis
Coronavirus Infections - virology
Coronavirus Nucleocapsid Proteins
Coronaviruses
COVID-19
Data Accuracy
Deoxyribonucleic acid
Diagnosis
Diagnostic equipment (Medical)
Diagnostic systems
Disease control
Disease transmission
DNA
DNA sequencers
DNA, Viral - genetics
Ebola virus
Emergency response
Engineering and Technology
Epidemics
Experiments
Fluorescence
Gene sequencing
Genetic testing
Humans
Influenza
Medicine and health sciences
N protein
Nucleic acids
Nucleocapsid Proteins - genetics
Nucleotide sequence
Pandemics
Phosphoproteins
Pneumonia, Viral - diagnosis
Pneumonia, Viral - virology
Point-of-Care Systems
Polymerase chain reaction
Polymerase Chain Reaction - economics
Polymerase Chain Reaction - instrumentation
Polymerase Chain Reaction - methods
Portable equipment
Power supply
Protocol
Reagents
Reproducibility of Results
Research and Analysis Methods
SARS-CoV-2
Sensitivity and Specificity
Severe acute respiratory syndrome
Severe acute respiratory syndrome coronavirus 2
Supervision
Viral diseases
Viruses
Zika virus
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Description
Summary:The coronavirus disease 2019 (COVID-19) pandemic has crudely demonstrated the need for massive and rapid diagnostics. By the first week of July, more than 10,000,000 positive cases of COVID-19 have been reported worldwide, although this number could be greatly underestimated. In the case of an epidemic emergency, the first line of response should be based on commercially available and validated resources. Here, we demonstrate the use of the miniPCR, a commercial compact and portable PCR device recently available on the market, in combination with a commercial well-plate reader as a diagnostic system for detecting genetic material of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causal agent of COVID-19. We used the miniPCR to detect and amplify SARS-CoV-2 DNA sequences using the sets of initiators recommended by the World Health Organization (WHO) for targeting three different regions that encode for the N protein. Prior to amplification, samples were combined with a DNA intercalating reagent (i.e., EvaGreen Dye). Sample fluorescence after amplification was then read using a commercial 96-well plate reader. This straightforward method allows the detection and amplification of SARS-CoV-2 nucleic acids in the range of ~625 to 2×105 DNA copies. The accuracy and simplicity of this diagnostics strategy may provide a cost-efficient and reliable alternative for COVID-19 pandemic testing, particularly in underdeveloped regions where RT-QPCR instrument availability may be limited. The portability, ease of use, and reproducibility of the miniPCR makes it a reliable alternative for deployment in point-of-care SARS-CoV-2 detection efforts during pandemics.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0237418