TY - JOUR
T1 - RNA polymerization actuating nucleic acid membrane (RANAM)-based biosensing for universal RNA virus detection
AU - Kim, Dajeong
AU - Han, Sangwoo
AU - Ji, Yoonbin
AU - Youn, Heejeong
AU - Kim, Hyejin
AU - Ko, Ohsung
AU - Lee, Jong Bum
N1 - Publisher Copyright:
© 2021
PY - 2022/3/1
Y1 - 2022/3/1
N2 - The coronavirus disease (COVID-19) pandemic has shown the importance of early disease diagnosis in preventing further infection and mortality. Despite major advances in the development of highly precise and rapid detection approaches, the time-consuming process of designing a virus-specific diagnostic kit has been a limiting factor in the early management of the pandemic. Here, we propose an RNA polymerase activity-sensing strategy utilizing an RNA polymerization actuating nucleic acid membrane (RANAM) partially metallized with gold for colorimetric RNA virus detection. Following RANAM-templated amplification of newly synthesized RNA, the presence of the RNA polymerase was determined by visualization of the inhibition of an oxidation/reduction (redox) reaction between 3,3′,5,5′-tetramethylbenzidine (TMB) and blocked Au3+. As a proof of concept, a viral RNA-dependent RNA polymerase (RdRP), which is found in various RNA virus-infected cells, was chosen as a target molecule. With this novel RANAM biosensor, as little as 10 min of RdRP incubation could significantly reduce the colorimetric signal. Further development into an easy-to-use prototype kit in viral infection diagnosis detected RdRP present at levels even as low as 100 aM. Color formation based on the presence of RdRP could be simply and clearly confirmed through smartphone-assisted color imaging of the prototype kit. This study provides a non-PCR-based RNA virus detection including its variants using RdRP-mediated polymerization.
AB - The coronavirus disease (COVID-19) pandemic has shown the importance of early disease diagnosis in preventing further infection and mortality. Despite major advances in the development of highly precise and rapid detection approaches, the time-consuming process of designing a virus-specific diagnostic kit has been a limiting factor in the early management of the pandemic. Here, we propose an RNA polymerase activity-sensing strategy utilizing an RNA polymerization actuating nucleic acid membrane (RANAM) partially metallized with gold for colorimetric RNA virus detection. Following RANAM-templated amplification of newly synthesized RNA, the presence of the RNA polymerase was determined by visualization of the inhibition of an oxidation/reduction (redox) reaction between 3,3′,5,5′-tetramethylbenzidine (TMB) and blocked Au3+. As a proof of concept, a viral RNA-dependent RNA polymerase (RdRP), which is found in various RNA virus-infected cells, was chosen as a target molecule. With this novel RANAM biosensor, as little as 10 min of RdRP incubation could significantly reduce the colorimetric signal. Further development into an easy-to-use prototype kit in viral infection diagnosis detected RdRP present at levels even as low as 100 aM. Color formation based on the presence of RdRP could be simply and clearly confirmed through smartphone-assisted color imaging of the prototype kit. This study provides a non-PCR-based RNA virus detection including its variants using RdRP-mediated polymerization.
KW - Nucleic acid membrane
KW - Nucleic acid metallization
KW - RNA virus detection
KW - RNA-dependent RNA polymerase
KW - SARS-CoV-2
UR - http://www.scopus.com/inward/record.url?scp=85121205795&partnerID=8YFLogxK
U2 - 10.1016/j.bios.2021.113880
DO - 10.1016/j.bios.2021.113880
M3 - Article
C2 - 34915215
AN - SCOPUS:85121205795
SN - 0956-5663
VL - 199
JO - Biosensors and Bioelectronics
JF - Biosensors and Bioelectronics
M1 - 113880
ER -