SENSORS AND ACTUATORS B-CHEMICAL
·
2022
·
IF 9.221
Ting Chen,Pengfei Liu,Huanxiang Wang,Yue Su,Sheng Li,Shimeng Ma,Xuan Xu,Jie Wen,Zhen Zou
Accurate detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is of great importance to control the COVID-19 pandemic. The gold standard assays for COVID-19 diagnostics are mainly based on separately detecting open reading frame 1ab (ORF1ab) and nucleoprotein (N) genes by RT-PCR. However, the current approaches often obtain false positive-misdiagnose caused by cross-contamination or undesired amplification. To address this issue, herein, we proposed a dumbbell-type triplex molecular switch (DTMS)-based, logic-gated strategy for high-fidelity SARS-CoV-2 RNA detection. The DTMS consists of a triple-helical stem region and two-loop regions for recognizing the ORF1ab and N genes of SARS-CoV-2. Only when the ORF1ab and N gene are concurrent, DTMS experiences a structural rearrangement, thus, bringing the two pyrenes into spacer proximity and leading to a new signal readout. This strategy allows detecting SARS-CoV-2 RNA with a detection limit of 1.3 nM, independent of nucleic acid amplification, holding great potential as an indicator probe for screening of COVID-19 and other population-wide epidemics.
研究领域:
分子生物学
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病毒学
·
诊断学
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DOI
PubMed
高影响因子
TALANTA
·
2023
·
IF 6.1
Yaqi Hu,Fang Bao,Siyuan Fu,Shiyuan Feng,Jinfeng Miao,Peng Miao,Yuanyuan Xu
Coronaviruses are highly infectious and pose a serious threat to human and animal healths. In this work, a facile electrochemical method based on Exonuclease III (Exo III) catalyzed digestion and silver deposition is developed for coronavirus RNA analysis. A magnetic separation procedure is performed to specifically identify target sequence and release single-stranded DNA modified gold nanoparticles (AuNPs). The nanoparticles can thus be immobilized at a screen-printed electrode and catalyze silver deposition for signal readout. This method allows sensitive analysis of PEDV and SARS-CoV-2 RNAs in the concentration range from 1 to 1000 nM with the limits of detection as low as 0.47 nM and 0.17 nM, respectively. Good specificities are demonstrated. Thus, the proposed method may have great potential use in the applications of coronaviruses analysis.
研究领域:
生物医学工程
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分子生物学
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DOI
PubMed
高影响因子
Journal of Materials Chemistry B
·
2018
·Vol. 6
·期 26
·4380-4384
·
IF 4.776
Guanghan Li,Yong Liu,Jie Niu,Meishan Pei,Weiying Lin
RNA plays a central role in controlling cellular functions. Research of the content and distribution of RNA in living cells is of great significance to both biochemistry and biomedicine. However, ratiometric fluorescent probes for the detection of RNA in the cytoplasm and nucleoli are still rarely reported. We herein present the first example of a novel ratiometric fluorescent composite nanomaterial by using graphene quantum dots (GQDs) and a fluorescent probe molecule for the sensitive and selective detection of RNA. HVC-6 was selected as the detection group. The fluorescence was excited at 365 nm and the fluorescence emission at 470 and 610 nm increased gradually with the addition of RNA. The fluorescence intensity ratio of I610/I470 displayed a linear response to RNA. Furthermore, the developed nanomaterial HVC-6@GQDs showed potential for utilization as a fluorescent RNA probe in living cells.
研究领域:
生物化学
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生物医学
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纳米材料科学
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DOI
PubMed
高影响因子
PLoS One
·
2023
·Vol. 18
·期 3
·e0268917
·
IF 3.7
Chen Zhang,Qingrong Qu,Yuming Yao,Xiaobo Fan,Guoqiu Wu
The hybridization chain reaction (HCR) is widely used for biosensing. However, HCR does not provide the required sensitivity. In this study, we reported a method to improve the sensitivity of HCR by dampening the cascade amplification. First, we designed a biosensor based on HCR, and an initiator DNA was used to trigger the cascade amplification. Optimization of the reaction was then performed, and the results showed that the limit of detection (LOD) for the initiator DNA was about 2.5 nM. Second, we designed a series of inhibitory DNAs to dampen the HCR cascade amplification, and DNA dampeners (50 nM) were applied in the presence of the DNA initiator (50 nM). One of the DNA dampeners (D5) showed the best inhibitory efficiency of greater than 80%. This was further applied at concentrations ranging from 0 nM to 10 nM to prohibit the HCR amplification caused by a 2.5 nM initiator DNA (the limit of detection for this initiator DNA). The results showed that 0.156 nM of D5 could significantly inhibit the signal amplification (p<0.05). Additionally, the limit of detection for the dampener D5 was 16 times lower than that for the initiator DNA. Based on this detection method, we achieved a detection limit as low as 0.625 nM for HCV-RNAs. In summary, we developed a novel method with improved sensitivity to detect the target designed to prohibit the HCR cascade. Overall, this method could be used to qualitatively detect the presence of single-stranded DNA/RNA.
研究领域:
分子生物学
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医学检验
·
生物化学
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DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2015
·
IF 5.636
Jingpu Zhang,Chao Li,Xiao Zhi,Gabriel Alfranca Ramón,Yanlei Liu,Chunlei Zhang,Fei Pan,Daxiang Cui
MicroRNA (miRNA) biomarkers display great potential for cancer diagnosis and prognosis. The development of rapid and specific methods for miRNA detection has become a hotspot. Herein, hairpin DNA-templated silver nanoclusters (AgNCs/HpDNA) were prepared and integrated into strand-displacement amplification (SDA) as a novel beacon for miRNA detection. The light-up platform was established based on guanine (G)-rich fluorescence enhancement that essentially converted the excitation/emission pair of AgNCs/HpDNAs from a shorter wavelength to a longer wavelength, and then achieved fluorescent enhancement at longer wavelength. On the basis of the validation of the method, the single and duplex detection were conducted in two plasma biomarkers (miR-16-5p and miR-19b-3p) for the diagnosis of gastric cancer. The probe (AgNCs/RED 16(7s)C) utilized for miR-16-5p detection adopted a better conformation with high specificity to recognize single-base mismatches by producing dramatically opposite signals (increase or decrease at 580 nm ex/640 nm em) while the probe (AgNCs/GRE 19b(5s)C) for miR-19b-3p generated dual signals (increase at 490 nm ex/570 nm em and decrease at 430 nm ex/530 nm em) with bright fluorescence in one reaction during the amplification, but unexpectedly was partially digested. This is for the first time to allow the generation of enhanced fluorescent AgNCs and the target recognition integrated into a single process, which offers great opportunity for specific miRNA detection in an easy and rapid way.
研究领域:
生物化学
·
分子生物学
·
癌症研究
·
纳米技术
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF ELECTROANALYTICAL CHEMISTRY
·
2020
·
IF 3.807
Ziheng Hu,Biying Zhao,Ping Miao,Xialing Hou,Feifei Xing,Yingying Chen,Lingyan Feng
Herein, we present one three-way junction (TWJ) DNA based electrochemical biosensor for sensitive microRNAs detection with locked nucleic acid (LNA) as capture probe and enzyme-free redox cycling signal amplification strategy. LNA owns higher binding affinity and thermal stability compared to normal DNA sequence . In the presence of target microRNA (miRNA-21), novel TWJ structure can be specially triggered with the assistant of methylene blue (MB)-tagged DNA signal probe, thus producing an enhanced electrochemical current. Furthermore, a chemical reductant, tris (2-carboxyethyl) phosphine (TCEP), is adopted to lead a relayed electron transport between solution TCEP and TWJ-linked MB tag and amplified whole signal intensity. Based on the selectivity from target-induced LNA/DNA TWJ structure and enzyme-free MB cycling amplification strategy, the designed electrochemical biosensor exhibits sensitive miRNA-21 detection, with a linear range from 100 aM to 100 pM and a low detection limit of 77 aM. In addition, it can successfully discriminate the differences between the miRNA family members, presenting a promising response to the direct determination of target miRNA-21 in real biological fluids. Our work provides an alternative LNA-based electrochemical biosensor way in the area of miRNA diagnostics and clinical analysis without complicated enzyme/nanomaterials-assisted signal amplification.
研究领域:
生物化学
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分子诊断
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分析化学
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产品关联
DOI
高影响因子
ACS Sensors
·
2025
·
IF 9.1
Yu Luo,Bicheng Zhu,Congcong Zhu,Penghui Lai,John Taylor,Claire Honney,Ashley Nutsford,Chaofeng Ma,Hailong Chen,Kean C. Aw,Rui Wu,Erasmus Smit,Peikai Zhang,Jadranka Travas-Sejdic
Herein, a novel and simple electrospray (ES) printing technique was developed for the fabrication of ultrathin graphene layers with precisely controlled nanometer-scale thickness, where graphene oxide (GO) was electrosprayed on wafers and subsequently chemically reduced into reduced GO (rGO). Utilizing that technique, we prepared ultrathin rGO in-plane graphene field-effect transistor (GFET)-based biosensors coupled with a portable prototype measuring system for point-of-care detection of pathogens. We illustrate the use of such prepared GFETs to detect COVID-19, using the SARS-CoV-2 nucleocapsid protein antigen (N-protein) and genomic viral RNA as detection targets. The electrosprayed and chemically reduced rGO films enhance the molecular detection in GFET sensors through significant local gating effects. The device detects the N-protein from the SARS-CoV-2 Omicron variant in a culture medium with an LOD of 1.44 PFU/mL and in clinical oropharyngeal samples with an LOD of 45 genome copies/mL in 5 min. It also successfully detects viral RNA in oropharyngeal swabs within 10 min. The GFET sensor responses were further analyzed using our proprietary wireless, miniaturized, and portable FET analyzer, coupled with a smartphone detecting app. Altogether, we present low-cost and mass-producible GFETs with high-quality graphene channels, enabling a portable, efficient, and accurate solution for point-of-care pathogen detection and in clinical testing. This technology has the potential to become a crucial tool in preventing future global epidemic outbreaks.
研究领域:
生物传感器
·
医学诊断
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纳米技术
·
病毒学
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产品关联
DOI
PubMed
高影响因子
ADVANCED FUNCTIONAL MATERIALS
·
2024
·
IF 18.5
Yuteng Tang,Yuanhang Xiang,Yu Yang,Yuyuan Zhang,Binqi Wei,Xiaojie Qin,Min Fang,Qiqin Wang,Xinchun Li,Fan Yang
The unmet on-site clinical diagnostics are increasingly demanding portable bioassays, but compromised by sensitivity and throughput. Herein, a nanostructured bubbles-enhanced fluorescence (nBEF) assay is reported by interfacing with smartphone imaging system to enable portable, ultrasensitive, and specific miRNA detection in parallel. In nBEF, the high-curvature bubbles allow physically enhancing optical propagation, chemically enhancing fluorescence, assembling more DNA probes, undergoing faster self-suspension and more efficient self-aggregation fluorescence enhancement relative to their smooth counterparts. By coupling with bubble-driven signal amplification by a plug-and-play DNA concatemer, and with a smartphone imaging system, this nBEF assay achieves a portable and ultrasensitive fluorescent assay of miRNA at the femtomolar level with one-base resolution in a single test, and is capable of monitoring drug-induced miRNA dynamic expression and accurate cancer diagnosis (AUC = 1) through profiling 40 plasma samples after RNA extraction. This assay further integrates an R-value analysis in RGB module with the smartphone to realize automated, on-site miRNA readout. Such a high-performance nBEF assay provides a low-cost, portable, and highly sensitive tool to enable accurate clinical diagnosis.
研究领域:
生物化学
·
分子诊断
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医学工程
·
临床检验
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产品关联
DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2020
·
IF 7.1
Hui Kong,Wei Zhang,Jia Yao,Chao Li,Renfei Lu,Zhen Guo,Jinze Li,Chuanyu Li,Yingxue Li,Chiyu Zhang,Lianqun Zhou
Development of a simple, sensitive and specific method for HIV-1 assay is demanded. In some emergency situations, a portable reverse transcription loop-mediated isothermal amplification (RT-LAMP) platform is necessary. A novel hydrogen ion-selective RT-LAMP (H + -RT-LAMP) sensor was developed for highly sensitive HIV-1 using hydrogen ion concentration as a signal producing compound. The H + -RT-LAMP sensor was based on screen-printed with carbon-doped graphene conductive layer, carbon nanotubes (CNTs) solid electrolyte layer, and reference electrode KCl functional layer, which provided superior electrical signals, high sensitivity and stability for HIV-1 detection. The rate of hydrogen ion production was related to the viral load (VL) during the amplification of HIV-1. The sensitive HIV-1 RNA detection range was achieved from 0 copies/μL to 10 5 copies/μL. The proposed H + -RT-LAMP sensor was applied to detect HIV-1 RNA in real human serum. 57 samples were detected and satisfactory results were obtained. Compared with commercial instruments, the detection results of H + -RT-LAMP sensor demonstrated a perfect concordance rate of 96.5 %, and the limit of detection (LOD) for the clinical samples was 10 copies per tube. Time to get the result is from 10 min to 25 min. Therefore, the proposed H + -RT-LAMP sensor was particularly suitable as a point-of-care device for the on-site detection of HIV-1.
研究领域:
分子生物学
·
医学检验
·
生物传感器
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产品关联
DOI
高影响因子
International Journal of Nanomedicine
·
2023
·
IF 8
Dawei Deng,Yang Li,Jianpeng Xue,Jie Wang,Guanhua Ai,Xin Li,Yueqing Gu
Messenger RNA (mRNA), a single-strand ribonucleic acid with functional gene information is usually abnormally expressed in cancer cells and has become a promising biomarker for the study of tumor progress. Hairpin DNA-coated gold nanoparticle (hDAuNP) beacon containing a bare gold nanoparticle (AuNP) as fluorescence quencher and thiol-terminated fluorescently labeled stem–loop–stem oligonucleotide sequences attached by Au–S bond is currently a new nanoscale biodiagnostic platform capable of mRNA detection, in which the design of the loop region sequence is crucial for hybridizing with the target mRNA. Hence, in this study, to improve the sensitivity and selectivity of hDAuNP beacon simultaneously, the loop region of hairpin DNA was screened by bioinformatics strategy. Here, signal transducer and activator of transcription 5b (STAT5b) mRNA was selected and used as a practical example. The results from the combined characterizations using optical techniques, flow cytometry assay, and cell microscopic imaging showed that after optimization, the as-prepared hDAuNP beacon had higher selectivity and sensitivity for the detection of STAT5b mRNA in living cells, as compared with our previous beacon. Thus, the bioinformatics method may be a promising new strategy for assisting in the designing of the hDAuNP beacon, extending its application in the detection of mRNA expression and the resultant mRNA-based biological processes and disease pathogenesis.
研究领域:
纳米技术
·
分子生物学
·
生物信息学
·
癌症研究
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产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2018
·
IF 5.667
Lei Wang,Qi Xia,Ruiyuan Liu,Jinqing Qu
A red fluorescent probe, PFB, is designed and synthesized, which presents several remarkable features including a large Stokes shift (85 nm), long emission wavelength, high photostability and good biocompatibility. PFB also displays the solvent-dependent effect, which is verified by the density functional theory (DFT) calculations. Additionally, PFB can act as a fluorescent probe for cellular imaging with high uptake efficiency and highly sensitive detection toward RNA under low optimum concentration (1 μM), which enables it to achieve cancer cell tracing in vitro and tumor growth monitoring in vivo . The results of real-time tracking experiment reveal that PFB can be traced in stained HepG-2 cells for 11 generations and monitor tumor growth for 18 days. This work highlights the potential application of PFB as a promising alternative to the commercial RNA fluorescent probes.
研究领域:
生物化学
·
分子生物学
·
医学影像学
·
肿瘤学
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产品关联
DOI
高影响因子
ANALYTICA CHIMICA ACTA
·
2025
·
IF 6
Hong Zhang,Xuekun Bai,Chenxi Wang,Wei Pang,Jiaxin Zhang,Junhuai Song,Huanying Zhou,Zhixian Gao,Bin Zheng
Background Biorhythmic played an important role in physical health. Biorhythmic disorders led to various health issue, including cardiovascular diseases and metabolic disorders. Early diagnosis of biorhythmic disorders was crucial in slowing the progression of disease. Therefore, it was important to develop technologies for detecting biorhythmic and their disorder markers. This study selected HMGB1(High mobility group box 1), BMAL1 (Brain and Muscle Aryl hydrocarbon receptor nuclear translocator-like 1), and MICU (mitochondrial calcium uptake 1) mRNA for detection. Results CRISPR/Cas12a/SERS (surface-enhanced Raman spectroscopy) ultrasensitive biosensing platform was developed for the detection of biorhythmic markers. The biosensor was partly an ultrabright Raman signal reporter (on) of a novel satellite structure formed by gold/silver bimetallic nanoparticles. This novel satellite structure utilizes the potential difference between gold and silver nanoparticles to promote the transfer of electrons from the core structure to the AuNPs and MBA (4-mercaptobenzoic acid) molecules, while enhancing the electromagnetic field distribution. Compared with the Raman signal of the satellite structure composed of Au/Au bimetallic, the Raman signal of the satellite structure in this study is increased by about 10 times. CRISPR/Cas12a recognized the target molecule and activated trans -cleavage activity, cleaving the DNA/RNA hairpin structure and releasing RNA 2∗ . RNA 2∗ caused the satellite structure to disperse, significantly attenuating the ultra-strong Raman signal (off), thereby enabling quantitative detection of the target nucleic acid. The LOD for three targets were of less than 1 fM. Significance In summary, the CRISPR/Cas12a/SERS biosensing platform enables mRNA detection without preamplification. This biosensing platform can detect target molecules at the fM level, achieving ultra-sensitive detection. In addition, the satellite structure exhibits long-term stability, providing a theoretical basis for the practical application of this biosensing platform.
研究领域:
生物传感器
·
分子诊断学
·
纳米技术
·
CRISPR技术
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RNA检测
·
生物医学工程
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产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）