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TALANTA · 2025 · IF 6.1

Advancement of doxorubicin monitoring with a DNA fragmentation strategy for SYBR Green I-based aptamer biosensors

Luke Wei,Yuxin Hu,Jiamin Wu,Jing Mao,Tianqin Yin,Jieqiong Qiu

Doxorubicin (DOX), a widely utilized chemotherapeutic antibiotic, increasingly threatens environmental and public health due to its persistence in marine ecosystems, bioaccumulation in organisms, and contamination of agricultural systems. In response to the urgent demand for effective DOX detection, we have developed a label-free fluorescent aptamer biosensor (FAB) that employs SYBR Green I (SG I)-mediated signal amplification. A key innovation in this design is the strategic fragmentation of the 41-mer complementary strand (CP) of the DOX aptamer into three 11-mer fragments. It significantly mitigates nonspecific SG I binding interference caused by the longer ssDNA CP, enhancing the fluorescence signal ratio (F dsDNA /F ssDNA ) from 4.6-fold to 12.6-fold. Upon DOX introduction, competitive binding between DOX, SG I, and DNA further suppresses background noise, yielding an exceptional fluorescence quenching ratio (F 0 /F) of 52.8. The optimized FAB demonstrates ultra-sensitive DOX detection, with a detection limit of 1.41 nM and a linear response between 1.41 nM and 300 nM. The recoveries of DOX spiked into pond water and drinking water samples ranged from 95.1% to 101.8%, with relative standard deviations (RSDs) below 3.46%, demonstrating negligible matrix effects and confirming the FAB method’s robustness for environmental monitoring applications. This strategy synergistically enhances signal transduction efficiency, and strengthens detection reliability, all while maintaining operational simplicity. As a result, this FAB for DOX detection holds transformative potential, poised to revolutionize biosensor technology in the fields of food safety, environmental monitoring, and biosecurity.

研究领域: 生物传感器 · 环境科学 · 分析化学 · 分子生物学 · 生物化学 · 纳米技术 · 公共卫生 · 海洋生态学
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JOURNAL OF FLUORESCENCE · 2024 · IF 2.6

Label-Free Detection of Programmed Death-Ligand 1 for Prognosis Using a Truncated Aptamer and SYBR Green I

Li Ling,Li Zheng-Ling,Wang Wei,Long Xin-Xin,Liu Ping,Wang Guo-Tian,Sun Shi-Qing,Du Rong-Lian

The rapid and accurate detection of programmed death-ligand 1 (PD-L1) expression is of great value in the diagnosis and treatment of tumors. ELISA-based traditional method is the gold standard for protein detection, but there are still some shortcomings, especially the antigen-antibody dependence, greatly increased the detection time and cost. This work constructed a label-free fluorescent probe for rapid and sensitive detection of PD-L1 using a truncated aptamer as recognition molecules and double-stranded DNA specific dyes (SYBR Green I) as signal units. After a series of optimization conditions, this probe has good detection capability for PD-L1 in buffer solution with the detection limit as low as 0.68 ng/mL. Due to the specific recognition ability of aptamer and target, this method also has good selectivity for PD-L1 detection. The recovery of PD-L1 in human serum samples ranges from 86.20 to 96.36%. Compared with other methods, this strategy does not need to be marked, and does not need other complex design and purification process, but simple operation process and strong anti-interference ability. The whole detection process can be completed within 20 min and has good application prospect. This work will provide reference for drug dosage and prognosis evaluation of specific tumor therapy.

CAS / MDL: 50-81-7 51-61-6
研究领域: 癌症生物学 · 分子诊断 · 生物技术
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ANALYST · 2013 ·Vol. 138 ·期 17 ·4737-4740 · IF 3.969

An enzyme-free and label-free assay for copper(II) ion detection based on self-assembled DNA concatamers and Sybr Green I

Chenchen Ge,Junhua Chen,Wei Wu,Zhiyuan Fang,Lingbo Chen,Qi Liu,Lin Wang,Xuerong Xing,Lingwen Zeng

An enzyme-free and label-free fluorescence turn on biosensor for amplified copper(II) ion (Cu2+) detection has been constructed based on self-assembled DNA concatamers and Sybr Green I. This assay is simple, inexpensive and sensitive, enabling quantitative detection of as low as 12.8 pM Cu2+.

研究领域: 分析化学 · 生物传感器 · 纳米技术
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FOOD CHEMISTRY · 2019 · IF 5.399

A dichromatic label-free aptasensor for sulfadimethoxine detection in fish and water based on AuNPs color and fluorescent dyeing of double-stranded DNA with SYBR Green I

Xiang-Xiu Chen,Zheng-Zhong Lin,Cheng-Yi Hong,Qiu-Hong Yao,Zhi-Yong Huang

A dichromatic label-free aptasensor was described for sulfadimethoxine (SDM) detection. Compared with the binding of SDM-aptamer to SDM, the higher affinity of aptamer to cDNA may result in the hybridization of dsDNA. In the presence of SDM, the aptamer specifically binds to SDM, leading to a blue color of AuNPs in deposit and fluorescence at 530 nm in supernatant after adding cDNA and SGI. With no target of SDM, AuNPs protected with the aptamer re-disperse in PBS with a red color, and no fluorescence occurs in supernatant. Based on the principle, SDM can be quantitatively detected through both fluorescent emission and AuNPs color changes with recoveries ranging from 99.2% to 102.0% for fish and from 99.5% to 100.5% for water samples. An analytical linear range of 2–300 ng mL −1 was achieved with the detection limits of 3.41 ng mL −1 for water and 4.41 ng g −1 for fish samples (3 σ , n  = 9).

CAS / MDL: 122-11-2 72-14-0
研究领域: 分析化学 · 环境科学 · 食品安全
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FOOD CHEMISTRY · 2022 · IF 9.231

Aptamer truncation strategy assisted by molecular docking and sensitive detection of T-2 toxin using SYBR Green I as a signal amplifier

Pengfei Ma,Hua Ye,Hualin Guo,Xiaoyuan Ma,Lin Yue,Zhouping Wang

Herein, for the first time, a novel truncation strategy for aptamers’ sequences based on molecular docking was reported for the first time. T-2 toxin and AFB 1 aptamers were employed as models and fluorescence polarization was used to measure the affinity of aptamers. The T-2 toxin aptamer (T40) with 40 bases and the AFB 1 aptamer with 32 bases were successfully obtained with increased affinities compared with original aptamers. Meanwhile, circular dichroism, FAM-BHQ1 dual labeled T40, enzyme digestion, and molecular dynamics simulation were performed to investigate the binding mechanism between aptamer T40 and T-2 toxin. Finally, based on MnO 2 nanosheet and the fluorescence amplification effect of SYBR Green I, simplified and rapid fluorescence detection of T-2 toxin was achieved with a detection range of 0.03 nM to 30 nM, indicating the great potential of aptamer in practical detection applications.

研究领域: 分析化学 · 分子生物学 · 毒理学 · 纳米技术
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MICROCHEMICAL JOURNAL · 2018 · IF 2.746

Label-free fluorescent aptasensor of Cd2+ detection based on the conformational switching of aptamer probe and SYBR green I

Bin Zhou,Xin-Yi Yang,Yong-Sheng Wang,Jie-Can Yi,Zhen Zeng,Huan Zhang,Ya-Ting Chen,Xi-Jiang Hu,Qing-Li Suo

A label-free fluorescence aptasensor based on conformational switching of aptamer induced by Cd 2+ was established for the good selective determination of Cd 2+ . The aptamer could act as a recognition element for Cd 2+ . SYBR green I (SG), as a signal reporter, can bind to the small groove of double-stranded DNA (dsDNA) constituted the aptamer and the complementary strand, establishing the dsDNA-SG complex and generating high fluorescence intensity . In the presence of Cd 2+ , the specific recognition and binging of aptamers preferentially with Cd 2+ induce free of complementary strands from dsDNA and conformational switching of aptamer from dsDNA to a stem-loop structure, fluorescence intensity of system decrease dramatically. Under the optimum experiment parameters, the aptasensor shows a good linear range from 1.12 μg L −1 to 224.82 μg L −1 and the excellent detection limit as low as 0.34 μg L −1 . The relative standard deviations (RSD) for determination of Cd 2+ (n = 8) were lower than 4%, and the recoveries were in the range of 98.57% ~ 102.49%. The use of fluorescence aptasensor has good selectivity and does not require any sample treatment or target preconcentration for the detection of Cd 2+ in real samples.

CAS / MDL: 77-86-1
研究领域: 分析化学 · 环境科学 · 生物化学
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MICROCHEMICAL JOURNAL · 2024 · IF 4.9

Label-free photosensitization colorimetric detection of microRNA by utilizing cascade toehold assembly amplification

Yizhi Zhou,Qiang Liu

The sensitive and label-free detection of microRNA (miRNA) is crucial for the comprehension of miRNA-related pathological process, such as glioma. Here, we utilized a SYBR Green I (SG) catalytic photosensitization colorimetric reaction in conjunction with target recognition-induced exonuclease-iii (Exo-iii) assisted cascade toehold assembly amplification to achieve highly sensitive and label-free miRNA detection. In this research, the photosensitized generation of 1 O 2 by SG could directly oxidize 3,3′,5,5′-tetramethylbenzidine (TMB) without the need for H 2 O 2 oxidation or enzyme catalysis, which simplifies the experimental procedures and renders the method highly stable. Furthermore, the Exo-iii enhances the target recognition induced catalytic chain assembly, thereby forming cascade signal amplification and providing the method with a significantly higher level of sensitivity than conventional colorimetric miRNA detection methods. Based on this, the proposed method exhibits a low limit of detection of 2.1 fM for miRNA-212 detection. Taking the merit of the high sensitivity, portability, and simplicity, the proposed method can serve as a highly effective nucleic acid detection tool and can facilitate the advancement of biosensors for point-of-care testing (POCT) and clinical disease diagnosis.

CAS / MDL: 54827-17-7
研究领域: 生物化学 · 分子诊断 · 医学检测
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BIOSENSORS & BIOELECTRONICS · 2016 · IF 7.476

Binding induced colocalization activated hybridization chain reaction on the surface of magnetic nanobead for sensitive detection of adenosine

Chunjing Feng,Zhun Hou,Wei Jiang,Lihong Sang,Lei Wang

Herein, a sensitive and enzyme-free assay for adenosine detection has been developed on the basis of binding induced colocalization activated hybridization chain reaction (HCR) strategy on the surface of magnetic nanobead . First, the recognition probe was fabricated and divided into two parts: the Apt-1 that composed a part of adenosine aptamer and toehold domain, and the Apt-2 that consisted of another part of adenosine aptamer and branch migration domain. The Apt-1 was immobilized on a streptavidin-magnetic nanobead (streptavidin-MNBs) that played the roles of enrichment and separation. Then the recognition event of adenosine could bring the two parts of aptamer together and induce the colocalization of toehold domain and branch migration domain, which could serve as an integrated initiator to trigger the HCR, producing a long nicked double-stranded polymer. Finally, the intercalating dye SYBR Green I was inserted into the polymer, generating an enhanced fluorescence signal. In this strategy, the initiator was divided into two parts and could be suppressed effectively in the absence of adenosine. Utilizing the separated function, the spontaneous hybridization of H1 and H2 could be avoided, and a low background could be acquired. Moreover, through the double amplification of HCR and multimolecules binding of SYBR Green I, highly sensitive and enzyme-free detection were achieved. The detection limit for adenosine detection was 2.0×10 −7   mol/L, which was comparable or superior to the previous aptasensors . Importantly, adenosine analysis in human urines has been performed, and this strategy could significantly distinguish the adenosine content in normal human urines and cancer patient urines, suggesting that this proposed assay will become a reliable and sensitive adenosine detection method in early clinical diagnosis and medical research.

CAS / MDL: 51-61-6
研究领域: 生物化学 · 医学诊断 · 分析化学
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AQUACULTURE · 2025 · IF 3.9

A DNAzyme biosensor for highly sensitive and rapid detection of decapod iridovirus 1

Shuai Zhang,Siying Li,Huili Tian,Ming'ao Wang,Mingsheng Lyu,Shujun Wang

Decapod Iridescent Virus 1 (DIV1) leads to high mortality rates in shrimp farming, significantly impacting the aquaculture industry economically. Currently, there are no effective treatments for controlling DIV1 infections, and management primarily relies on preventative measures. Rapid and precise diagnosis of DIV1 is crucial. In this study, we identified two DNA enzymes, MCP-1 and MCP-8, that can specifically recognize DIV1 via in vitro selection. The detection limit of MCP-1 was 6.89 copies/μL of DIV1. We developed a DNAzyme-based “turn off” biosensor for quick detection of DIV1, which is both highly specific and sensitive, by integrating SYBR Green I to enhance the signal. The detection limit of biosensor could reach 2.01 copies/μL of DIV1. Compared to conventional detection methods, this sensor delivers results in 5 min without the need for complex procedures, greatly streamlining the detection process.

研究领域: 分子生物学 · 水产养殖学 · 病毒学
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MICROCHIMICA ACTA · 2018 ·Vol. 185 ·期 2 ·1-7 · IF 5.705

Fluorometric determination of nucleic acids based on the use of polydopamine nanotubes and target-induced strand displacement amplification

Ge Jia,Bai Dong-Mei,-Geng Xin,Hu Ya-Lei,Cai Qi-Yong,Xing Ke,Zhang Lin,Li Zhao-Hui

The authors describe a fluorometric method for the quantitation of nucleic acids by combining (a) cycled strand displacement amplification, (b) the unique features of the DNA probe SYBR Green, and (c) polydopamine nanotubes. SYBR Green undergoes strong fluorescence enhancement upon intercalation into double-stranded DNA (dsDNA). The polydopamine nanotubes selectively adsorb single-stranded DNA (ssDNA) and molecular beacons. In the absence of target DNA, the molecular beacon, primer and SYBR Green are adsorbed on the surface of polydopamine nanotubes. This results in quenching of the fluorescence of SYBR Green, typically measured at excitation/emission wavelengths of 488/518 nm. Upon addition of analyte (target DNA) and polymerase, the stem of the molecular beacon is opened so that it can bind to the primer. This triggers target strand displacement polymerization, during which dsDNA is synthesized. The hybridized target is then displaced due to the strand displacement activity of the polymerase. The displaced target hybridizes with another molecular beacon. This triggers the next round of polymerization. Consequently, a large amount of dsDNA is formed which is detected by addition of SYBR Green. Thus, sensitive and selective fluorometric detection is realized. The fluorescent sensing strategy shows very good analytical performances towards DNA detection, such as a wide linear range from 0.05 to 25 nM with a low limit of detection of 20 pM. Schematic of a fluorometric strategy for highly sensitive and selective determination of nucleic acids by combining strand displacement amplification and the unique features of SYBR Green I (SG) and polydopamine nanotubes.

研究领域: 分析化学 · 分子生物学 · 纳米技术
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MICROCHEMICAL JOURNAL · 2025 · IF 5.1

A label-free electrochemical biosensor for detection of uranyl ions in water based on nucleic acid dye-inducing turn-off of photosensitization

Mei Xiang,Chengyan Yin,Qiulian Chen,Guonian Zhu,Zhaoxia Chen,Hailiang Li,Ruth Antwi-Baah,Jiangwen Tang,Kunhong Yang,Shuxia Xu

A DNAzyme-based photocatalytic electrochemical biosensor was developed for the label-free detection of uranyl   ions (UO 2 2+ ) in water. In this sensing platform, SYBR Green (SG) molecules were embedded in double-stranded DNA (dsDNA), which was cleaved via formed singlet oxygen ( 1 O 2 ) during photosensitization, resulting in a significant decrease in the impedance signal of [Fe (CN) 6 ] 3−/4− . In the presence of UO 2 2+ , the substrate strand was cleaved by the target UO 2 2+ , releasing the SG molecule from the dsDNA, and the photosensitive reaction was turned off. Therefore, UO 2 2+ can be detected based on changes in [Fe(CN) 6 ] 3−/4− electrochemical signals. Such a sensing strategy allowed the detection of UO 2 2+ in the concentration range of 1–1000 nM, with a limit of detection (LOD) of 1.3 × 10 −10 M. The LOD meets the maximum allowable contamination level (1.3 × 10 −7 M) of UO 2 2+ in drinking water set by the US Environmental Protection Agency (EPA). The proposed sensor has been successfully applied for detection of UO 2 2+ in spiked actual water samples with recoveries in the range of 88.0 %–103 %, comparable to those obtained by ICP-MS. The method, which combines photocatalysis with an electrochemical biosensor, significantly improved the sensitivity for UO 2 2+ detection without the need of labeling and additional signal amplification steps. It is also a promising strategy for rapid monitoring of UO 2 2+ in environmental water samples.

研究领域: 环境科学 · 分析化学 · 生物传感技术
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COLLOIDS AND SURFACES A-PHYSICOCHEMICAL AND ENGINEERING ASPECTS · 2020 · IF 3.99

Biomacromolecular fluorescent nanoparticles co-assembled by bovine serum albumin and DNA segments for living cell imaging

Si Sun,Qian-Ru Xiao,Cui-Fang Ye,Yong Jiang

Fluorescent nanoparticles have been widely used in the field of biomedical engineering because they are usually biocompatible, cost-effective and homogeneously uniform. In this study, we describe a biomacromolecular fluorescent nanoparticle prepared with a facile disolvating process using bovine serum albumin, DNA segments and SYBR Green I. The obtained co-assemblies, termed [email protected] nanoparticles, had a narrow and relatively uniform size distribution with an average size of about 122 nm and the PDI value 0.087. Cellular uptake results demonstrated well internalization of [email protected] nanoparticles by the Michigan Cancer Foundation-7 (MCF-7) cells. Together with the fluorescent tag of SYBR Green I molecules, [email protected] nanoparticles accumulated in the cell cytoplasm after cellular internalization enabled the MCF-7 cells for fluorescent imaging. This study provided a new strategy for constructing biomacromolecular fluorescent nanoparticles for applications in biomedical engineering and biomedical imaging.

CAS / MDL: 98-10-2
研究领域: 生物医学工程 · 分子生物学
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