Combined recombinase polymerase amplification/rkDNA-graphene oxide probing system for detection of SARS-CoV-2.
Anal Chim Acta
; 1158: 338390, 2021 May 08.
Article
in English
| MEDLINE | ID: covidwho-1141547
ABSTRACT
The development of rapid, highly sensitive, and selective methods for the diagnosis of infection by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) should help to prevent the spread of this pandemic virus. In this study, we combined recombinase polymerase amplification (RPA), as a means of isothermal DNA amplification, with an rkDNA-graphene oxide (GO) probe system to allow the rapid detection of SARS-CoV-2 with high sensitivity and selectivity. We used in situ enzymatic synthesis to prepare an rkDNA probe that was complementary to an RPA-amplified sequence of the target N-gene of SARS-CoV-2. The fluorescence of this rkDNA was perfectly quenched in the presence of GO. When the quenched rkDNA-GO system was added to the RPA-amplified sequence of the target SARS-CoV-2, the fluorescence recovered dramatically. The combined RPA/rkDNA-GO system exhibited extremely high selectivity (discrimination factor 17.2) and sensitivity (LOD = 6.0 aM) for the detection of SARS-CoV-2. The total processing time was only 1.6 h. This combined RPA/rkDNA-GO system appears to be a very efficient and simple method for the point-of-care detection of SARS-CoV-2.
Keywords
Full text:
Available
Collection:
International databases
Database:
MEDLINE
Main subject:
Nucleic Acid Amplification Techniques
/
SARS-CoV-2
/
COVID-19
/
Graphite
Type of study:
Diagnostic study
Limits:
Humans
Language:
English
Journal:
Anal Chim Acta
Year:
2021
Document Type:
Article
Affiliation country:
J.aca.2021.338390
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