TY - JOUR
T1 - Interference Reduction Biosensing Strategy for Highly Sensitive microRNA Detection
AU - Chen, Feng
AU - Li, Guodong
AU - Wu, Chun
AU - Wang, Ling
AU - Ko, Chung Nga
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); Guangdong Basic and Applied Basic Research Foundation (grant no. 2021A1515110338); the Science and Technology Development Fund, Macau SAR, China (file no. 0007/2020/A1); and SKL-QRCM(UM)-2020-2022; the University of Macau, China (MYRG2020-00017-ICMS).
Publisher Copyright:
© 2022 American Chemical Society
PY - 2022/3/15
Y1 - 2022/3/15
N2 - MicroRNAs are potential biomarkers for human cancers and other diseases due to their roles as post-transcriptional regulators for gene expression. However, the detection of miRNAs by conventional methods such as RT-qPCR, in situ hybridization, northern blot-based platforms, and next-generation sequencing is complicated by short length, low abundance, high sequence homology, and susceptibility to degradation of miRNAs. In this study, we developed a nicking endonuclease-mediated interference reduction rolling circle amplification (NEM-IR-RCA) strategy for the ultrasensitive and highly specific detection of miRNA-21. This method exploits the advantages of the optical properties of long-lived iridium(III) probes, in conjunction with time-resolved emission spectroscopy (TRES) and exponential rolling circle amplification (E-RCA). Under the NEM-IR-RCA-based signal enhancement processes, the limit of detection of miRNA-21 was down to 0.0095 fM with a linear range from 0.05 to 100 fM, which is comparable with the conventional RT-qPCR. Unlike RT-qPCR, the strategy was performed at a lower and constant temperature without heating/cooling cycles and reverse transcription. The strategy could clearly discriminate between matched and mismatched targets, demonstrating high specificity. Moreover, the potential application of this method was demonstrated in cancer cells and mouse serum samples, showing good agreement with RT-qPCR results. Apart from miRNA-21 detection, this platform could be also adapted for detecting other miRNAs, such as let-7a and miRNA-22, indicating its excellent potential for biomedical research and clinical diagnostics.
AB - MicroRNAs are potential biomarkers for human cancers and other diseases due to their roles as post-transcriptional regulators for gene expression. However, the detection of miRNAs by conventional methods such as RT-qPCR, in situ hybridization, northern blot-based platforms, and next-generation sequencing is complicated by short length, low abundance, high sequence homology, and susceptibility to degradation of miRNAs. In this study, we developed a nicking endonuclease-mediated interference reduction rolling circle amplification (NEM-IR-RCA) strategy for the ultrasensitive and highly specific detection of miRNA-21. This method exploits the advantages of the optical properties of long-lived iridium(III) probes, in conjunction with time-resolved emission spectroscopy (TRES) and exponential rolling circle amplification (E-RCA). Under the NEM-IR-RCA-based signal enhancement processes, the limit of detection of miRNA-21 was down to 0.0095 fM with a linear range from 0.05 to 100 fM, which is comparable with the conventional RT-qPCR. Unlike RT-qPCR, the strategy was performed at a lower and constant temperature without heating/cooling cycles and reverse transcription. The strategy could clearly discriminate between matched and mismatched targets, demonstrating high specificity. Moreover, the potential application of this method was demonstrated in cancer cells and mouse serum samples, showing good agreement with RT-qPCR results. Apart from miRNA-21 detection, this platform could be also adapted for detecting other miRNAs, such as let-7a and miRNA-22, indicating its excellent potential for biomedical research and clinical diagnostics.
UR - http://www.scopus.com/inward/record.url?scp=85126513937&partnerID=8YFLogxK
U2 - 10.1021/acs.analchem.2c00138
DO - 10.1021/acs.analchem.2c00138
M3 - Journal article
AN - SCOPUS:85126513937
SN - 0003-2700
VL - 94
SP - 4513
EP - 4521
JO - Analytical Chemistry
JF - Analytical Chemistry
IS - 10
ER -