MICROCHIMICA ACTA
·
2025
·Vol. 192
·期 3
·1-10
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IF 5.3
Liu Jianghua,Wang Yu,Zhang Xinyue,Huang Mingquan,Li Guoliang
A novel direct lysis method combined with amplification-free CRISPR/Cas12a-SERS genosensor was for the first time developed to rapidly and sensitively identify meat adulteration. Notably, polystyrene (PS) microspheres, with distinct shrinking and swelling properties, were dexterously employed to encapsulate biological-silent Raman reporter 4-mercaptobenzonitrile (4-MBN) and act as a controlled-release signal probe. Target DNA activated the trans -cleavage activity of CRISPR/Cas12a towards ssDNA linked with PS microsphere to liberate the signal probe, which was able to release numerous Raman reporters after treatment with THF solution, resulting in high signal amplification. Through this platform, trace target DNA was deftly transformed into a sensitive Raman signal and could be on-site determined through a portable Raman equipment. Under optimized conditions, this strategy displayed good linearity in the range 1–450 ng/μL ( R 2 = 0.9943) and favorable sensitivity with limit of detection as low as 0.23 ng/μL without any pre-amplification. Moreover, it exhibited good applicability to on-site identification of commercial meat samples in complicated food matrix. In addition, DNA extraction by direct lysis and amplification-free detection realized ultrafast meat adulteration determination within 35 min from sampling to result. This method possessed great potential in rapid and on-site accurate determination of meat authenticity. Graphical abstract
研究领域:
食品科学
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分子生物学
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分析化学
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产品关联
DOI
PubMed
高影响因子
Nanoscale
·
2013
·Vol. 5
·期 13
·5833-5840
·
IF 6.233
Li-qiang Luo,Zhao Zhang,Ya-ping Ding,Dong-mei Deng,Xiao-li Zhu,Zhen-xin Wang
In this work, we proposed a novel simple protocol for preparing 1-aminopyrene/graphene (ApG) hybrids for fabricating label-free electrochemical impedance genosensor. Graphene, with the structure of a single-atom-thick sheet of sp2-bonded carbon atoms, was anchored to 1-aminopyrene (1-Ap) with the pyrenyl group via π-stacking interaction. The morphology, conductivity, and interaction of ApG hybrids were characterized by transmission electron microscopy (TEM), cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), UV-visible (UV-vis) and fluorescence spectra. The amino-substituted oligonucleotide probe was conjugated to 1-Ap by the cross-linker glutaraldehyde. The DNA hybridization reaction of oligonucleotide probe with target DNA was monitored by EIS. Under optimum conditions, the proposed biosensor exhibited high sensitivity and a low detection limit for detecting the complementary oligonucleotide. The target oligonucleotide could be quantified in a wide range of 1.0 × 10−12 to 1.0 × 10−8 M with good linearity (R = 0.9900) and low detection limit of 4.5 × 10−13 M (S/N = 3).
研究领域:
纳米材料
·
生物传感器
·
电化学
·
遗传学
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DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2025
·
IF 8.5
Shahab Uddin,Sun Shao Yan,Wei-Zhong Zhang,Qian-Wen Zhao,Jia-Xu Chen,Zhaorong Yue,Yang Li,Xin Wang,Hongyu Li
Periodontitis, a progressive inflammatory condition, causes periodontal tissue destruction, pain, chewing difficulties, and tooth loss, primarily due to pathogenic oral biofilms dominated by low abundance periodontopathic bacteria. Accurate diagnosis is hindered by the limited bacterial presence in saliva, necessitating effective enrichment techniques. This study introduces novel amoxicillin-loaded chitosan-coated magnetic nanocarriers (AMX-CS-MNCs) designed for rapid capture, enrichment, and DNA extraction of Gram-positive and Gram-negative anaerobic periodontal pathogens, including Porphyromonas gingivalis and Parvimonas micra , from artificial and clinical saliva samples collected at Kangmei Dental Hospital, facilitating highly sensitive PCR-based detection. The chitosan coating enhances biocompatibility and dispersibility, while amoxicillin acts as a broad-spectrum targeting ligand. Comprehensive characterization using X-ray photoelectron spectroscopy (XPS), X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), zeta-potential analysis, dynamic light scattering (DLS), vibrating sample magnetometry (VSM), and energy-dispersive X-ray spectroscopy (EDS) confirms successful functionalization and drug loading. AMX-CS-MNCs demonstrate exceptional bacterial capture efficiency (>90 %) at low concentrations following preconcentration, integrating with PCR to achieve a limit of detection (LOD) of 10 CFU/mL—a 1000-fold improvement over conventional PCR. Validation with patient samples demonstrated robust DNA amplification, highlighting this platform's potential for early, sensitive, and selective periodontitis clinical diagnosis and targeted therapy.
研究领域:
纳米技术
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生物医学工程
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微生物学
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牙科医学
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分子诊断学
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药物递送
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材料科学
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DOI
PubMed
高影响因子
Alexandria Engineering Journal
·
2024
·
IF 6.2
Jia Wang,Juelan Ye,Gang Li
Human papillomavirus (HPV) is the primary cause of cervical cancer, a major global health concern affecting women worldwide. Early detection of high-risk HPV genotypes is crucial for timely intervention and reducing disease burden. However, current detection methods often lack sensitivity, speed, or cost-effectiveness, especially in resource-limited settings. This work addresses this critical need by developing an ultrasensitive electrochemical biosensing platform based on nitrogen-doped graphene paper electrodes for quantifying high-risk HPV16 DNA sequences. The N-graphene nanohybrid, fabricated via simple solvothermal treatment of biomass-derived graphene oxide, demonstrated significantly enhanced electrical and electrocatalytic properties compared to undoped graphene. DNA detection was achieved through characteristic oxidation signals of guanine bases, which displayed a wide linear dynamic range (0.5–100 μg/mL), low detection limit (75 pg/mL), and excellent reproducibility (relative standard deviation <3.5 %). Detailed spectroscopic and electrochemical analyses revealed the key roles of pyridinic nitrogen defects in improving interfacial charge transfer kinetics and mitigating biofouling issues. This synergistic combination of high electrical conductivity and versatile surface functionality paves the way for equipment-free, point-of-care genosensor devices tailored for quantitative biomarker screening in resource-constrained environments.
研究领域:
生物医学工程
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电化学
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分子生物学
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癌症研究
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纳米技术
·
即时诊断
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DOI
高影响因子
危险品化学品经营许可证（带存储）