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Detection and differentiation of respiratory syncytial virus subgroups A and B with colorimetric toehold switch sensors in a paper-based cell-free system
- Source :
- Biosensorsbioelectronics. 182
- Publication Year :
- 2021
-
Abstract
- Respiratory syncytial virus (RSV) infection is the most common clinical infectious disease threatening the safety of human life. Herein, we provided a sensitive and specific method for detection and differentiation of RSV subgroups A (RSVA) and B (RSVB) with colorimetric toehold switch sensors in a paper-based cell-free system. In this method, we applied the toehold switch, an RNA-based riboswitch, to regulate the translation level of β-galactosidase (lacZ) gene. In the presence of target trigger RNA, the toehold switch sensor was activated and the expressed LacZ hydrolyzed chromogenic substrates to produce a colorimetric result that can be observed directly with the naked eye in a cell-free system. In addition, nucleic acid sequence-based amplification (NASBA) was used to improve the sensitivity by amplifying target trigger RNAs. Under optimal conditions, our method produced a visible result for the detection of RSVA and RSVB with the detection limit of 52 aM and 91 aM, respectively. The cross-reaction of this method was validated with other closely related respiratory viruses, including human coronavirus HKU1 (HCoV-HKU1), and Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). Furthermore, we used the paper-based carrier material that allows stable storage of our detection elements and rapid detection outside laboratory. In conclusion, this method can sensitively and specifically differentiate RSVA and RSVB and generate a visible colorimetric result without specialized operators and sophisticated equipment. Based on these advantages above, this method serves as a simple and portable detector in resource-poor areas and point-of-care testing (POCT) scenarios.
- Subjects :
- Riboswitch
Biomedical Engineering
Biophysics
02 engineering and technology
Biosensing Techniques
01 natural sciences
Virus
Cell-free system
Betacoronavirus
Electrochemistry
Humans
Gene
biology
Cell-Free System
Chemistry
SARS-CoV-2
010401 analytical chemistry
Nucleic acid sequence
RNA
General Medicine
021001 nanoscience & nanotechnology
biology.organism_classification
NASBA
Virology
0104 chemical sciences
Respiratory Syncytial Virus, Human
RNA, Viral
Human coronavirus HKU1
Colorimetry
0210 nano-technology
Biotechnology
Subjects
Details
- ISSN :
- 18734235
- Volume :
- 182
- Database :
- OpenAIRE
- Journal :
- Biosensorsbioelectronics
- Accession number :
- edsair.doi.dedup.....969da8d7d526d6cdc573a1a96450bb0b