ANALYTICA CHIMICA ACTA
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2017
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IF 4.95
Cuiying Lin,Yajuan Guo,Mengmeng Zhao,Mi Sun,Fang Luo,Longhua Guo,Bin Qiu,Zhenyu Lin,Guonan Chen
Development of simple but sensitive biosensor for influenza detection is highly important in immediate and effective clinical treatment. In this study, a sensitive colorimetric immunosensor which combines the advantages of high selectivity of immunoassay and simplicity of colorimetric detection has been developed to detect influenza virus H5N1 based on enzyme-encapsulated liposome. Biotin-tagged liposome encapsulated with large amount of horseradish peroxidase (HRP) was firstly synthesized. In the presence of H5N1, H5N1 co-bound with the capture antibody and the biotinylated detection antibody to form sandwich immunocomplex . Subsequently, the HRP-encapsulated liposome was introduced to conjugate with the detection antibody through biotin-avidin-biotin linkage. Upon the addition of substrate (mixture of 3,3′,5,5′-tetramethylbenzidine (TMB) and H 2 O 2 ), the liposome was directly lysed to release large amount of HRP by TMB. The released HRP catalyzed the H 2 O 2 -mediated oxidation of TMB, resulting in color change of the system, which was observed by naked eyes or UV–vis spectra. The result showed that the absorption intensity enhanced with the increase of H5N1 concentration ranging from 0.1 to 4.0 ng/mL, and the detection limit was calculated to be 0.04 ng/mL. The sensitivity of the proposed biosensor is much higher than that of conventional enzyme-linked immunosorbent assay method. The proposed immunosensor is relatively simple, low-cost, sensitive, and selective without using any sophisticated instruments, therefore it may have a promising prospect for detecting targets in clinical medicine, food safety analysis, and environmental monitoring.
研究领域:
生物技术
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病毒学
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分析化学
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医学诊断
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DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
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2016
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IF 5.886
Cheng-Yu Li,Di Cao,Ya-Feng Kang,Yi Lin,Ran Cui,Dai-Wen Pang,Hong-Wu Tang
We present an analytical platform by combining near-infrared optical tweezers with two-photon excitation for fluorescence detection of H5N1 virus gene sequences. A heterogeneous enrichment strategy, which involved polystyrene (PS) microsphere and quantum dots (QDs), was adopted. The final hybrid-conjugate microspheres were prepared by a facile one-step hybridization procedure by using PS microspheres capturing target DNA and QDs tagging, respectively. Quantitative detection was achieved by the optical tweezers setup with a low-cost 1064 nm nanosecond pulse laser for both optical trapping and two-photon excitation for the same hybrid-conjugate microsphere. The detection limits for both neuraminidase (NA) gene sequences and hemagglutinin (HA) gene sequences are 16–19 pM with good selectivity for one-base mismatch, which is approximately 1 order of magnitude lower than the most existing fluorescence-based analysis method. Besides, because of the fact that only signal from the trapped particle is detected upon two-photon excitation, this approach showed extremely low background in fluorescence detection and was successfully applied to directly detect target DNA in human whole serum without any separation steps and the corresponding results are very close to that in buffer solution, indicating the strong anti-interference ability of this method. Therefore, it can be expected to be an emerging alternative for straightforward detecting target species in complex samples with a simple procedure and high-throughput.
研究领域:
生物技术
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分子生物学
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光学分析
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DOI
高影响因子
ANALYTICAL CHEMISTRY
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2018
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IF 6.042
Yang Wang,Qingyu Ruan,Zhi-Chao Lei,Shui-Chao Lin,Zhi Zhu,Leiji Zhou,Chaoyong Yang
Digital microfluidics (DMF) is a powerful platform for a broad range of applications, especially immunoassays having multiple steps, due to the advantages of low reagent consumption and high automatization. Surface enhanced Raman scattering (SERS) has been proven as an attractive method for highly sensitive and multiplex detection, because of its remarkable signal amplification and excellent spatial resolution. Here we propose a SERS-based immunoassay with DMF for rapid, automated, and sensitive detection of disease biomarkers. SERS tags labeled with Raman reporter 4-mercaptobenzoic acid (4-MBA) were synthesized with a core@shell nanostructure and showed strong signals, good uniformity, and high stability. A sandwich immunoassay was designed, in which magnetic beads coated with antibodies were used as solid support to capture antigens from samples to form a beads–antibody–antigen immunocomplex. By labeling the immunocomplex with a detection antibody-functionalized SERS tag, antigen can be sensitively detected through the strong SERS signal. The automation capability of DMF can greatly simplify the assay procedure while reducing the risk of exposure to hazardous samples. Quantitative detection of avian influenza virus H5N1 in buffer and human serum was implemented to demonstrate the utility of the DMF-SERS method. The DMF-SERS method shows excellent sensitivity (LOD of 74 pg/mL) and selectivity for H5N1 detection with less assay time (<1 h) and lower reagent consumption (∼30 μL) compared to the standard ELISA method. Therefore, this DMF-SERS method holds great potentials for automated and sensitive detection of a variety of infectious diseases.
研究领域:
分析化学
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生物工程
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生物医学工程
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病毒学
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微流控技术
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DOI
PubMed
高影响因子
JOURNAL OF MEDICINAL CHEMISTRY
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2014
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IF 5.48
Yuanchao Xie,Dongqing Xu,Bing Huang,Xiuli Ma,Wenbao Qi,Fangyuan Shi,Xinyong Liu,Yingjie Zhang,Wenfang Xu
To discover group-1-specific neuraminidase (NA) inhibitors that are especially involved in combating the H5N1 virus, two series of oseltamivir derivatives were designed and synthesized by targeting the 150-cavity. Among these, compound 20l was the most potent N1-selective inhibitor, with IC50 values of 0.0019, 0.0038, and 0.0067 μM against NAs from three H5N1 viruses. These values are better than those of oseltamivir carboxylate. Compound 32 was another potent N1-selective inhibitor that exhibited a 12-fold increase in activity against the H274Y mutant relative to oseltamivir carboxylate. Molecular docking studies revealed that the 150-cavity was an auxiliary binding site that may contribute to the high selectivity of these compounds. The present work is a significant breakthrough in the discovery of potent group-1-specific neuraminidase inhibitors, which may be further investigated for the treatment of infection by the H5N1 virus.
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DOI
PubMed
高影响因子
EUROPEAN JOURNAL OF MEDICINAL CHEMISTRY
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2012
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IF 3.346
Zhibo Zhu,Runming Li,Gaokeng Xiao,Zhipeng Chen,Jie Yang,Qiuhua Zhu,Shuwen Liu
We reported previously that a small molecule named CL-385319 could inhibit H5N1 influenza virus infection by targeting hemagglutinin , the envelope protein mediating virus entry. In the present study, a novel series of derivatives focused on the structural variation of CL-385319 were synthesized as specific inhibitors against the H5 subtype of influenza A viruses . These small molecules inhibited the low pH-induced conformational change of hemagglutinin, thereby blocking viral entry into host cells. Compound 1l was the most active inhibitor in this series with an IC 50 of 0.22 μM. The structure–activity relationships analysis of these compounds showed that the 3-fluoro-5-(trifluoromethyl)benzamide moiety was very important for activity, and the –F group was a better substituent group than –CF 3 group in the phenyl ring. The inhibitory activity was sensitive to the benzamide because the oxygen and hydrogen of the amide served as H-bond acceptor and donor, respectively.
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产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
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2024
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IF 8
Qiudi Xu,Ke Liu,Yu He,Lan Wang,Zefan Lu,Zhongxuan Liu,Tao Zhang
Digital microfluidics (DMF), as an emerging liquid-handling technology, is widely recognized as an ideal platform for enzyme-linked immunosorbent assay (ELISA). However, current DMF-based ELISA methods are highly dependent on magnetic beads (MBs), which usually involves tedious modification of MBs, complex peripherals for MBs control, and impairs the parallel processing capability of DMF. To address these issues, we herein developed a photopatterned reactive sites-integrated DMF chip (PRS-DMF) for MBs-free immunoassay. By taking advantage of click chemistry between N-(allyloxycarbonyloxy) succinimide and thiol-ene substrate, the preparation of PRS is very simple, and leaves the reactive N-hydroxysuccinimide (NHS) ester groups on the surface. This enables direct biomolecules immobilization without any surface modification/activation processes. Moreover, the patterned micropillar array can notably increase the specific surface area, and promote the liquid mixing and immunoreaction. As a consequence, highly sensitive and MBs-free ELISA can be readily achieved on PRS-DMF, with a proof-of-concept H5N1 assay having been successfully demonstrated herein, which offers a low limit of detection (LOD) of 147 pg/mL. This also avoids the use of complex magnetic peripherals, and thus, contributing to the instrument miniaturization into a very compact form. It is therefore reasonably expected that the proposed PRS-DMF may serve as a promising platform for a variety of biological and biomedical applications, especially in the field of point-of-care testing (POCT).
研究领域:
生物技术
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医学检验
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微流控技术
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免疫学
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产品关联
DOI
高影响因子
MICROCHEMICAL JOURNAL
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2026
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IF 5.1
Liang Gong,Lin Zhu,Wencheng Xu,Xiuzhi Shan,Miduo Tan,Yubin Gu,Juan Zhou,Sihua Liu
Influenza virus infections, particularly those caused by the Avian influenza virus, result in millions of deaths and substantial economic losses annually. Early diagnosis of the pathogen during the initial stages of an outbreak is pivotal for effective outbreak control. This study presents the development of an Exo III-mediated high sensitivity dual signal amplification (HSDSA) platform for detecting the H5N1 gene sequence. The detection mechanism is initiated by the hybridization of the target H5N1 DNA with hairpin DNA (Hp DNA) to form double-stranded structure. Hp DNA in this structure could be recognized and partial hydrolyzed by Exo III, releasing target DNA and free DNA. On one side, the target DNA enabled hybridize with other Hp DNA for circulate freely, achieving the first round of amplification. On the other hand, free DNA can hybridize with the DNA structure labeled on the gold nanoparticles, transforming the originally 3-terminal protruding double-stranded structure into a 3-terminal concave Exo III substrate structure. After triggering the Exo III recognition, the long chains (SH DNA) within dsDNA are hydrolyzed, and free DNA is released again to enter the next round of triggering for the second round of amplification. Meanwhile, another DNA labeled with carboxyfluorescein (FAM DNA) released from the hydrolyzed SH DNA, causing the fluorophore to detach from the AuNPs, the fluorescence signal is restored. This strategy significantly enhances detection sensitivity through double cyclic signal amplification mediated by Exo III, with a linear range from 0.08 nM to 15 nM and a detection limit of 54 pM for H5N1 DNA. The fluorescence sensor demonstrates robust applicability in detecting H5N1 in human serum samples, with recovery rates ranging from 98.75% to 105.83%. These results underscore the potential of the HSDSA platform for low-abundance molecular detection, disease diagnosis, and biomedicine applications.
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产品关联
DOI
高影响因子
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