TY - JOUR
T1 - A rapid and label-free DNA-based interference reduction nucleic acid amplification strategy for viral RNA detection
AU - Chen, Feng
AU - Li, Guodong
AU - Wu, Chun
AU - Wang, Wanhe
AU - Ma, Dik Lung
AU - Leung, Chung Hang
N1 - Funding Information:
This work is supported by Hong Kong Baptist University , the Health and Medical Research Fund, China ( HMRF / 14150561 ); the National Natural Science Foundation of China , China ( 22077109 and 21775131 ); the Science and Technology Development Fund, Macau SAR, China (File no. 0016/2020/A and 0007/2020/A1); SKL-QRCM(UM)-2020-2022; the University of Macau , China ( MYRG2019–00002–ICMS ).
Publisher Copyright:
© 2021 Elsevier B.V.
PY - 2022/2/15
Y1 - 2022/2/15
N2 - Common reference methods for COVID-19 diagnosis include thermal cycling amplification (e.g. RT-PCR) and isothermal amplification methods (e.g. LAMP and RPA). However, they may not be suitable for direct detection in environmental and biological samples due to background signal interference. Here, we report a rapid and label-free interference reduction nucleic acid amplification strategy (IR-NAAS) that exploits the advantages of luminescent iridium(III) probes, time-resolved emission spectroscopy (TRES) and multi-branch rolling circle amplification (mbRCA). Using IR-NAAS, we established a luminescence approach for diagnosing COVID-19 RNAs sequences RdRp, ORF1ab and N with a linear range of 0.06–6.0 × 105 copies/mL and a detection limit of down to 7.3 × 104 copies/mL. Moreover, the developed method was successfully applied to detect COVID-19 RNA sequences from various environmental and biological samples, such as domestic sewage, and mice urine, blood, feces, lung tissue, throat and nasal secretions. Apart from COVID-19 diagnosis, IR-NAAS was also demonstrated for detecting other RNA viruses, such as H1N1 and CVA10, indicating that this approach has great potential approach for routine preliminary viral detection.
AB - Common reference methods for COVID-19 diagnosis include thermal cycling amplification (e.g. RT-PCR) and isothermal amplification methods (e.g. LAMP and RPA). However, they may not be suitable for direct detection in environmental and biological samples due to background signal interference. Here, we report a rapid and label-free interference reduction nucleic acid amplification strategy (IR-NAAS) that exploits the advantages of luminescent iridium(III) probes, time-resolved emission spectroscopy (TRES) and multi-branch rolling circle amplification (mbRCA). Using IR-NAAS, we established a luminescence approach for diagnosing COVID-19 RNAs sequences RdRp, ORF1ab and N with a linear range of 0.06–6.0 × 105 copies/mL and a detection limit of down to 7.3 × 104 copies/mL. Moreover, the developed method was successfully applied to detect COVID-19 RNA sequences from various environmental and biological samples, such as domestic sewage, and mice urine, blood, feces, lung tissue, throat and nasal secretions. Apart from COVID-19 diagnosis, IR-NAAS was also demonstrated for detecting other RNA viruses, such as H1N1 and CVA10, indicating that this approach has great potential approach for routine preliminary viral detection.
KW - Coronavirus detection
KW - G-quadruplex
KW - Iridium(III) complex
KW - Rolling circle amplification
UR - http://www.scopus.com/inward/record.url?scp=85120177371&partnerID=8YFLogxK
U2 - 10.1016/j.bios.2021.113829
DO - 10.1016/j.bios.2021.113829
M3 - Journal article
C2 - 34840016
AN - SCOPUS:85120177371
SN - 0956-5663
VL - 198
JO - Biosensors and Bioelectronics
JF - Biosensors and Bioelectronics
M1 - 113829
ER -
