A Near-Infrared-Driven Signal Amplification Fluorescence Biosensor Based on Upconversion Beacon Probe for microRNA Detection

doi:10.19894/j.issn.1000-0518.230083

Chinese Journal of Applied Chemistry ›› 2024, Vol. 41 ›› Issue (1): 137-146.DOI: 10.19894/j.issn.1000-0518.230083

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A Near-Infrared-Driven Signal Amplification Fluorescence Biosensor Based on Upconversion Beacon Probe for microRNA Detection

Xue-Min ZHOU¹, Shu-Zhen LYU², Guo-Fang ZHANG¹, Zhu-Mei CUI¹(), Sai BI²()

^1.The Affiliated Hospital of Qingdao University，Qingdao 266061，China
^2.College of Chemistry and Chemical Engineering，Qingdao University，Qingdao 266071，China

Received:2023-03-31 Accepted:2023-05-31 Published:2024-01-01 Online:2024-01-30
Contact: Zhu-Mei CUI,Sai BI
About author:bisai11@126.com
cuizhumei1966@126.com
Supported by:
the Beijing Kanghua Foundation for the Development of Traditional Chinese and Western Medicine(KH-2021-LLZX-014);the National Natural Science Foundation of China(22076087);the Natural Science Fund for Distinguished Young Scholars of Shandong Province(ZR2020JQ08)

Abstract

Abstract:

In this work， a near-infrared light （NIR）-driven fluorescence biosensor is proposed based on the upconversion beacon probe and catalytic hairpin reaction （CHA） for the detection of microRNA. The target microRNA-21 （miRNA-21） can trigger the CHA reaction between the DNA hairpin H1 modified on magnetic beads （MBs） and the DNA hairpin H2 modified on upconversion nanoparticles （UCNPs）， resulting in the introduction of UCNPs on the surface of MBs accompanied by the cyclic signal amplification. Through magnetic separation， the UCNPs fixed on MBs are separated and generate upconversion luminescence （UCL） under the excitation of 808 nm NIR， which shows the advantages of good luminescence stability and negligible photodamage. In this biosensor， the linear range of miRNA-21 is 0.1 to 100 nmol/L with the detection limit of 11.3 pmol/L. In addition， this fluorescence biosensor achieves the detection of miRNA-21 in serum samples， showing good practicability.

Key words: Upconversion nanoparticles, Catalytic hairpin reaction, Fluorescence biosensor, MicroRNA, Near-infrared light

CLC Number:

O655

Xue-Min ZHOU, Shu-Zhen LYU, Guo-Fang ZHANG, Zhu-Mei CUI, Sai BI. A Near-Infrared-Driven Signal Amplification Fluorescence Biosensor Based on Upconversion Beacon Probe for microRNA Detection[J]. Chinese Journal of Applied Chemistry, 2024, 41(1): 137-146.

Figures/Tables 9

Fig.1 Schematic of fluorescence biosensor based on UCNPs and CHA amplification strategy for the detection of miRNA-21

Table 1 Sequences of oligonucleotides used in this study

Name	Sequence（5'-3'）
miRNA-21	UAG CUU AUC AGA CUG AUG UUG A
miRNA-155	UUA AUG CUA AUC GUG AUA GGG GU
H1	AGA CTG ATG TTG ACT TAG CTT ATC GAT CAA CAT CAG TCT GAT AAG CTA-NH₂
H2	ACT TAG CTT ATC AGA CTG ATG TTG ATC GAT AAG CTA AGT CAA CTA CA
Single-base mismatched miRNA-21	UAT CUU AUC AGA CUG AUG UUG A
Three-base mismatched miRNA-21	UAT CUA AUU AGA CUG AUG UUG A

Fig.2 （A） SEM image of MBs. TEM images of （B） NaYF4∶Yb3+，Tm3+ UCNPs and （C） NaYF4∶Yb3+，Tm3+@NaYF4 UCNPs. （D） XPS survey spectrum of NaYF4∶Yb3+，Tm3+@NaYF4 UCNPs. High-resolution XPS spectra of （E） Y3d and （F） F1s. TEM images of （G） OA-free UCNPs and （H） H2-UCNPs. （I） HRTEM and （J） EDS elemental mapping of H2-UCNPs. （K） Zeta potential of OA-free UCNPs and H2-UCNPs. （L） XRD spectrum of NaYF4∶Yb3+，Tm3+ UCNPs （a）， NaYF4∶Yb3+，Tm3+@NaYF4 UCNPs （b） and H2-UCNPs （c）. （M） UCL spectra of NaYF4∶Yb3+，Tm3+ UCNPs （a）， NaYF4∶Yb3+，Tm3+@NaYF4 UCNPs （b） and H2-UCNPs （c） under 808 nm NIR

Fig.3 Native PAGE for the characterization of CHA process. Lane 1： miRNA-21； lane 2： H1； lane 3： miRNA-21+H1； lane 4：miRNA-21+H1+H2； lane 5： H1+H2

Fig.4 （A） Optimization of H1 concentrations on MBs and （B） reaction time of CHA reaction

Fig.5 （A） UCL spectra toward miRNA-21 with different concentrations using the proposed NIR fluorescence biosensor. （B） Corresponding linear relationship between UCL intensity and logarithm of miRNA-21 concentration

Table 2 Comparison of performance of different sensing system

Methods	Linear range/（nmol·L^-１）	Detection limit/（pmol·L^-1）	Ref.
Fluorescence	0.1~16	93.8	［16］
Fluorescence	0.5~150	83.0	［31］
Fluorescence	0.1~300	67.0	［32］
Fluorescence	2.5~80	250.0	［33］
Electrochemistry	0.1~10⁴	83.0	［34］
Fluorescence	0.1~100	11.3	This work

Fig.6 （A） Specificity and （B） storage stability of the constructed NIR fluorescence biosensor

Table 3 Detection of miRNA-21 in human serum by NIR-driven fluorescence biosensor

Num	Added/（nmol·L^-1）	Measured value/（nmol·L^-1）	RSD/%（n=3）	Recovery/%
1	0	0.06	0.55	-
2	0.7	0.72	0.84	103.41
3	7	7.13	0.41	101.86
4	70	70.79	1.02	101.14

References 34

1	ALI SYDA Z， LANGDEN S， MUNKHZUL C， et al. Regulatory mechanism of microRNA expression in cancer［J］. Int J Mol Sci， 2020， 21（5）： 1723.
2	牛亚倩，刘芳，陈彻. 纳米生物传感器在肿瘤microRNA-21检测中的应用［J］. 中国生物制品学杂志， 2022， 35（6）： 759-762.
	NIU Y Q， LIU F， CHEN C. Application of nano-biosensors in detection of tumor microRNA-21［J］. Chin J Biol， 2022， 35（6）： 759-762
3	LI F， ZHOU Y， YIN H， et al. Recent advances on signal amplification strategies in photoelectrochemical sensing of microRNAs［J］. Biosens Bioelectron， 2020， 166： 112476.
4	VÁRALLYAY E， BURGYÁN J， HAVELDA Z. MicroRNA detection by northern blotting using locked nucleic acid probes［J］. Nat Protoc， 2008， 3（2）： 190-196.
5	SONG Y， XU Z， WANG F. Genetically encoded reporter genes for microRNA imaging in living cells and animals［J］. Mol Ther Nucleic Acids. 2020， 21： 555-567.
6	PETRICA L， PUSZTAI A M， VLAD M， et al. MiRNA expression is associated with clinical variables related to vascular remodeling in the kidney and the brain in type 2 diabetes mellitus patients［J］. Endocr Res， 2020， 45（2）： 119-130.
7	MEIS J E， KHANNA A. RNA amplification and cDNA synthesis for qRT-PCR directly from a single cell［J］. Nat Methods， 2009（6）： an12-an13.
8	于绍楠，任玲玲，任立群，等. 基于上转换纳米粒子-金纳米棒的荧光共振能量转移免疫分析法用于癌胚抗原检测［J］. 分析化学， 2022， 50（9）： 1299-1307.
	YU S N， REN L L， REN L Q， et al. Upconversion nanoparticles/gold nanorods-based fluorescence resonance energy transfer immunoassay for detection of carcinoembryonic antigen［J］. Chin J Anal Chem， 2022， 50（9）： 1299-1307.
9	HU Q， DUAN C， WU J， et al. Colorimetric and ratiometric chemosensor for visual detection of gaseous phosgene based on anthracene carboxyimide membrane［J］. Anal Chem， 2018， 90（14）： 8686-8691.
10	GUTIÉRREZ-G LVEZ L， GARCÍA-MENDIOLA T， GUTIÉRREZ-SÁNCHEZ C， et al. Carbon nanodot-based electrogenerated chemiluminescence biosensor for miRNA-21 detection［J］. Mikrochim Acta， 2021， 188（11）： 398.
11	LIU S T， CHEN J S， LIU X P， et al. A photoelectrochemical biosensor based on b-TiO₂/CdS∶Eu/Ti₃C₂ heterojunction for the ultrasensitive detection of miRNA-21［J］. Talanta， 2023， 253： 123601.
12	YANG Z， GUO Y， ZHOU J， et al. Ultrasensitive fluorescence detection and imaging of microRNA in cells based on a hyperbranched RCA-assisted multiposition SDR signal amplification strategy［J］. Anal Chem， 2022， 94（46）： 16237-16245.
13	ZHANG J， BAO X， ZHOU J， et al. A mitochondria-targeted turn-on fluorescent probe for the detection of glutathione in living cells［J］. Biosens Bioelectron， 2016， 85： 164-170.
14	ZHAO M， CHEN A Y， HUANG D， et al. MoS₂ quantum dots as new electrochemiluminescence emitters for ultrasensitive bioanalysis of lipopolysaccharide［J］. Anal Chem， 2017， 89（16）： 8335-8342.
15	HIMMELSTOß S F， HIRSCH T. A critical comparison of lanthanide based upconversion nanoparticles to fluorescent proteins， semiconductor quantum dots， and carbon dots for use in optical sensing and imaging［J］. Methods Appl Fluoresc， 2019， 7（2）： 022002.
16	ZHU Q Y， LI H， XU D K. Sensitive and enzyme-free fluorescence polarization detection for miRNA-21 based on decahedral sliver nanoparticles and strand displacement reaction［J］. RSC Adv， 2020， 10（29）： 17037-17044.
17	ZHAN Y， ZHANG R， GUO Y， et al. Recent advances in tumor biomarker detection by lanthanide upconversion nanoparticles［J］. J Mater Chem B， 2023， 11（4）： 755-771.
18	SHAO K， XIE W， LING Q， et al. Dumbbell-like upconversion nanoparticles synthesized by controlled epitaxial growth for light-heat-color tri-modal sensing of carcinoembryonic antigen［J］. Biosens Bioelectron， 2023， 228： 115186.
19	LIU Y， ZHAN S， SU X， et al. An optical strategy for detecting hypochlorite in vitro and cells with high selectivity and stability based on a lanthanide-doped upconversion probe［J］. RSC Adv， 2022， 12（49）： 31608-31616.
20	RONG Y， HASSAN M M， OUYANG Q， et al. Ratiometric upconversion fluorometric turn-off nanosensor for quantification of furfural in foods［J］. Sens Actuators B： Chem， 2022（350）： 130843.
21	龙禹同，万里，赵国杰. 催化发夹自组装技术用于miRNA检测的研究进展［J］. 生命科学研究， 2023， 27（1）： 86-94.
	LONG Y T， WAN L， ZHAO G J. Research progress of catalytic hairpin assembly technique for miRNA detection［J］. Life Sci Res， 2023， 27（1）： 86-94.
22	LIU J， ZHANG Y， XIE H， et al. Applications of catalytic hairpin assembly reaction in biosensing［J］. Small， 2019， 15（42）： e1902989.
23	SHEN J， LI T， WANG M， et al. Isothermal and enzyme-free microRNA assay based on catalytic hairpin assembly and rare earth element labeled probes［J］. Sens Actuators B： Chem， 2022， 357： 131364.
24	WANG Y X， WANG D X， WANG J， et al. DNA nanolantern-mediated catalytic hairpin assembly nanoamplifiers for simultaneous detection of multiple microRNAs［J］. Talanta， 2022， 236： 122846.
25	ZHANG X L， YIN Y， DU S M， et al. Dual 3D DNA nanomachine-mediated catalytic hairpin assembly for ultrasensitive detection of microRNA［J］. Anal Chem， 2021， 93（41）： 13952-13959.
26	ZHANG Y， ZHANG X， SITU B， et al. Rapid electrochemical biosensor for sensitive profiling of exosomal microRNA based on multifunctional DNA tetrahedron assisted catalytic hairpin assembly［J］. Biosens Bioelectron， 2021， 183： 113205.
27	KOSTIV U， FARKA Z， MICKERT M J， et al. Versatile bioconjugation strategies of PEG-modified upconversion nanoparticles for bioanalytical applications［J］. Biomacromolecules， 2020， 21（11）： 4502-4513.
28	LIU L， HUA R， ZHANG X， et al. Spectral identification and detection of curcumin based on lanthanide upconversion nanoparticles［J］. Appl Surf Sci， 2020， 525： 146566.
29	LUO Z， ZHANG L， ZENG R， et al. Near-infrared light-excited core-core-shell UCNP@Au@CdS upconversion nanospheres for ultrasensitive photoelectrochemical enzyme immunoassay［J］. Anal Chem， 2018， 90（15）： 9568-9575.
30	YAO C， TANG J， ZHU C， et al. A signal processor made from DNA assembly and upconversion nanoparticle for pharmacokinetic study［J］. Nano Today， 2022， 42： 101352.
31	YAO S， XIANG L， WANG L， et al. pH-responsive DNA hydrogels with ratiometric fluorescence for accurate detection of miRNA-21［J］. Anal Chim Acta， 2022， 1207： 339795.
32	ZHAO X， WANG S， ZOU R， et al. An enzyme-free probe based on G-triplex assisted by silver nanocluster pairs for sensitive detection of microRNA-21［J］. Mikrochim Acta， 2021， 188（2）： 55.
33	LU X， LI D， LUO Z， et al. A dual-functional fluorescent biosensor based on enzyme-involved catalytic hairpin assembly for the detection of APE1 and miRNA-21［J］. Analyst， 2022， 147（12）： 2834-2842.
34	KONG L Y， LV S Z， QIAO Z J， et al. Metal-organic framework nanoreactor-based electrochemical biosensor coupled with three-dimensional DNA walker for label-free detection of microRNA［J］. Biosens Bioelectron， 2022， 207： 114188.

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A Near-Infrared-Driven Signal Amplification Fluorescence Biosensor Based on Upconversion Beacon Probe for microRNA Detection

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