BIOSENSORS & BIOELECTRONICS
·
2013
·
IF 5.437
Wenjiao Zhou,Xue Gong,Yun Xiang,Ruo Yuan,Yaqin Chai
Visual detections have attracted great research attentions recently due to their convenient monitoring of the target analytes without using any advanced instruments. However, achieving visual detection of trace amounts of biomolecules with PCR-like sensitivity remains a major challenge. In current work, we describe a new quadratic signal amplification strategy for sensitive visual detection of HIV DNA biomarkers based on exonuclease III (Exo III)-assisted DNA recycling amplification and DNAzymes. The presence of the target HIV DNA leads to two independent and simultaneous DNA recycling processes to achieve quadratic signal amplification with the assistance of Exo III. This quadratic signal amplification results in catalytic cleavage of the G-quadruplex sequence-locked hairpin probes to release numerous active G-quadruplex sequences, which further associate with hemin to form DNAzymes and cause significantly intensified color change for sensitive and visual detection of HIV DNA down to 2.5 pM. The proposed visual detection method employs un-modified hairpin DNA as probes, avoids using any complex and expensive instruments for signal transduction and is essentially simple. This method also shows single-base mismatch discrimination capability as well. All these features make our developed DNA detection method holds great potential for visual monitoring of various DNA biomarkers at ultralow levels with careful and proper probe designs.
研究领域:
分子生物学
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2024
·
IF 5.7
Mengmeng Wu,Bing Yang,Lu Shi,Qiaorong Tang,Jing Wang,Wei Liu,Baoxin Li,Yan Jin
Background The human immunodeficiency virus (HIV) remains a major worldwide health problem. Nowadays, many methods have been developed for quantitative detecting human immunodeficiency virus DNA (HIV-DNA), such as fluorescence and colorimetry. However, these methods still have the disadvantages of being expensive and requiring professional technicians. Early diagnosis of pathogens is increasingly dependent on portable instruments and simple point-of-care testing (POCT). Therefore, it is meaningful and necessary to develop portable and cheap methods for detecting disease markers. Results In this work, a label-free chemiluminescence (CL) method was developed for detecting HIV-DNA via a handheld luminometer. To achieve label-free target detection, the CL catalyst, G-triplex-hemin DNAzyme (G3-hemin DNAzyme), was in-situ assembled in the presence of HIV-DNA. For improving sensitivity, HIV-DNA induced the cyclic strand displacement reaction (SDR), which can form three G3-hemin DNAzymes in one cycle. So, the chemiluminescence reaction between luminol and H 2 O 2 was highly effectively catalyzed, and the CL intensity was linearly related with the concentration of HIV-DNA in the range of 0.05–10 nM with a detection limit of 29.0 pM. Due to the high specificity of hairpin DNA, single-base mismatch can be discriminated, which ensured the specific detection of HIV-DNA. Significance In-situ formation of G3-hemin DNAzyme led to label-free and selective detection without complex synthesis and functionalization. Therefore, it offers a cheap, selective, sensitive and portable method for detecting disease-related genes, which is promising for POCT of clinical diagnosis in resource-limited settings.
研究领域:
生物医学
·
分子诊断
·
临床检验
✓ 已验证
产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2023
·
IF 8.4
Bing Yang,Lu Shi,Qiaorong Tang,Wei Liu,Baoxin Li,Chaoyong Yang,Yan Jin
Automated and sensitive detection of nucleic acid is crucial for reliable and safe disease diagnostics because the minimum manual operation can effectively avoid sample contamination and infection risk. Herein, a sensitive, label-free and automated method was developed for detecting trace DNA by integrating digital microfluidics (DMF) with chemiluminescence (CL) detection. DMF can automatically manipulate discrete droplets. Taking advantage of sensitive and simple CL detection, DMF-CL integration can automatically and sensitively detect HIV DNA via the preset pathway. To further improve the sensitivity, on-chip exponential amplification reaction (EXPAR) was explored to produce large quantities of double-stranded DNA (dsDNA) in the presence of HIV DNA. Then, SYBR Green I (SG) intercalated into dsDNA to generate singlet oxygen upon light irradiation , which in situ oxidized potassium ferrocyanide into potassium ferricyanide . Finally, the CL reaction of potassium ferricyanide and luminol ensured the label-free, reliable and specific detection of HIV DNA in the range of 10 pM to 2 nM. Saving time-consuming and laborious manual operations, the DMF-CL integration greatly reduced the reagent consumption and avoided the contamination of DNA sample as well. Therefore, DMF-CL offers a promising and useful technology for developing automated and reliable biosensing and bioanalysis.
研究领域:
生物医学工程
·
分析化学
·
分子生物学
·
诊断学
✓ 已验证
产品关联
DOI
高影响因子
International Journal of Electrochemical Science
·
2023
·
IF 1.5
Mingyi Ma,Li He,Xiaoxue Shi,Yanchao Wang,Hong Hai,Xiaoping Wei
This paper presents a novel strategy for ultrasensitive detection of human immunodeficiency virus (HIV) DNA by an electrochemical biosensor. By the shearing cycle of the target DNA (T-DNA), a large amount of complimentary sequence probes (CS probe DNA) are obtained. Then, the Zr 4+ functional probe DNA containing many branched DNAs is captured. Next, the initiator α-bromophenylacetic acid is connected to the branched chains of probe DNA by esterification reaction. Numerous electroactive monomers, namely, ferrocenylmethyl methacrylate (FMMA), are produced at the end of the branched chains by electrochemically mediated atomic transfer radical polymerization reaction. The signal of FMMA is greatly enhanced due to a large number of branched chain-loaded probe clusters containing numerous FMMA. T-DNA can be detected with a linear range of 0.1 fM–10 nM with a detection limit of 0.037 fM. The sensor is highly sensitive and selective. It has been applied to the detection of HIV DNA in human serum samples, and it has potential application prospects in the early clinical diagnosis of AIDS.
研究领域:
生物技术
·
分析化学
·
病毒学
·
电化学
✓ 已验证
产品关联
DOI
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2018
·
IF 8.173
Xilei Deng,Cong Wang,Ya Gao,Jingwen Li,Wei Wen,Xiuhua Zhang,Shengfu Wang
Up to now, the colloidal gold labeling immunochromatographic test strip is a mature and applicable technology. However, different from the conventional gold nanoparticle , quantum dot (QD) possesses larger specific surface area and better biocompatibility. So, as a novel nanomaterial , QD is capable of assembling more biomolecule which could enhance the sensitivity and accuracy of strips by rationality. Besides, strand displacement amplification was drawn into our test strips in this paper, this assumption made HIV-DNA recycling many times and converting it to plentiful QD-dsDNA (double-stranded deoxyribonucleic acid), where after these nano-structures would be captured by test zone. Meanwhile, the suggested scheme eliminated the hook effect owing to the target drop out of the incorporation on test zone, and any nucleotide sequence or substance which has aptamers can work as the target, such as carcinoembryonic antigen or mycotoxin . This assay realized the detection limit of as low as 0.76 pM (S/N = 3) and the detection range of 1 pM to 10 nM. In the end, we made use of this fluorescent lateral flow assay strips with great reproducibility for detecting HIV-DNA in human serum, that attested this method could be applied to practical application prospectively.
研究领域:
生物技术
·
分子生物学
·
纳米技术
·
病毒学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Sensors and Actuators Reports
·
2026
·
IF 7.6
Li-Kang Yin,Rui Huang,Ze-Lin Wang,Can Yang,Xue Mi,Han-Ying Zhan,Zhi-Qi Zhang
Human Immunodeficiency Virus (HIV) compromises the human immune system and remains one of the deadliest infectious diseases globally. Nucleic acid testing is a powerful tool for the early detection of HIV infection, capable of real-time HIV DNA detection without the limitation of a window period. In this study, a DNA nanostructure-based biosensor that leveraged interactions between functionalized gold nanoparticles (AuNPs) and three-dimensional (3D) DNA walkers were developed for ultrasensitive fluorescence detection of HIV DNA. The biosensor comprised blocked-walking particles (BPs) with locked DNAzyme and substrate particles (SPs) with quenched carboxyfluorescein (FAM) fluorophores. Upon encountering HIV DNA, the blocking strands hybridized more strongly with the HIV DNA, exposing the DNAzyme, which subsequently cleaved FAM on the SPs from the AuNPs and restoring fluorescence. As all substrate strands on SPs were cleaved, the BPs interacted with new SPs, significantly amplifying the fluorescent signal. The fluorescence intensity showed a linear correlation with HIV DNA concentration ranging from 0.02 to 6.00 nM, with a detection limit of 12.8 pM. The performance of biosensor was evaluated using spiked serum samples, demonstrating good precision and recovery rates. The DNA walker biosensor leveraged particle-to-particle interactions for rapid signal amplification, offering great potential for sensitive biosensors.
✓ 已验证
产品关联
DOI
高影响因子
JOURNAL OF ANALYTICAL ATOMIC SPECTROMETRY
·
2014
·Vol. 30
·期 4
·888-894
·
IF 3.396
Jing Hu,Xiandeng Hou,Peng Wu
Hydride generation atomic fluorescence spectrometry (HG-AFS) has been widely used for highly sensitive determination of more than 10 elements that are capable of hydride generation in geological, environmental and biological samples, but its bioanalytical potential has not been well-exploited yet. With CdS quantum dots (QDs) as both the signal labels for DNA sandwiching and analytes for HG-AFS (acid-released Cd2+ from QDs), here we proposed an ultrasensitive indirect method for HIV DNA analysis by HG-AFS. After a conventional sandwich-type hybridization reaction between capture DNA, target DNA and signal DNA labelled with CdS QD assembled silica microspheres, nitric acid was added to dissolve QDs and release Cd2+ from the QD-labels, which were then detected by HG-AFS. This efficient signal amplification strategy, coupled with the highly sensitive HG-AFS detection, gave rise to an impressive limit of detection of 0.8 aM and a wide dynamic concentration range (1 aM to 1 fM). This HG-AFS-based DNA biosensing platform holds great promise in future tumor marker detection and early cancerous diseases diagnosis.
研究领域:
生物分析化学
·
分子生物学
✓ 已验证
产品关联
DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2021
·
IF 7.46
Kai-Ren Zhao,Li Wang,Peng-Fei Liu,Xiao-Min Hang,Hui-Yi Wang,Shu-Ying Ye,Zhi-Jun Liu,Guo-Xi Liang
Herein, we report a “on-off” signal-switchable electrochemiluminescence (ECL) biosensor for simple, sensitive and quantitative detection of two kinds of virus genes, human immunodeficiency virus (HIV) and human papilloma virus (HPV-16) DNAs, by the integration of spherical nucleic acid (SNA) with CRISPR/Cas12a. The concept behind this approach is “on” signal based on a sandwich type assay after ECL luminophores of carbon dots (CDs) labeled SNA incubated with the biosensor in the presence of HIV DNA; and “off” signal upon the biosensor incubation with assistant DNA (HIV DNA sequence), CDs-labeled SNA and Cas12a/crRNA/HPV-16 DNA ternary complex relying on the collaterally cleavage of Cas12a. The cross-interference between HIV and HPV-16 DNAs can be avoided by adding Cas12a effector to the system or not. Using this strategy, HIV and HPV-16 DNAs are successfully quantitatively detection with detection limits of 30 fM and 0.32 pM (S/N = 3), respectively. Importantly, this system does not rely on multiplex signal reporting molecules, thus simplifying the detection system and avoiding the likelihood of cross interferences presented in the assay. Furthermore, multiplex detection of HIV/HPV-16 DNAs in human serum samples are successfully achieved within 2 h without additional nucleic acid amplification step. As demonstrated, the proposed biosensor has the potential for multiplex detection in clinical diagnosis.
研究领域:
生物传感技术
·
分子诊断学
·
病毒学
✓ 已验证
产品关联
DOI
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2015
·
IF 6.409
Zhongdan Yan,Ning Gan,Huairong Zhang,De Wang,Li Qiao,Yuting Cao,Tianhua Li,Futao Hu
A novel sandwich-hybridization assay for simultaneous electrochemical detection of multiple DNA targets related to human immune deficiency virus (HIV) and tuberculosis (TB) was developed based on the different quantum dots-PowerVision TM polymer nanotracers. The polymer nanotracers were respectively fabricated by immobilizing SH-labeled oligonucleotides (s-HIV or s-TB), which can partially hybrid with virus DNA (HIV or TB), on gold nanoparticles (Au NPs) and then modified with PowerVision TM (PV) polymer-encapsulated quantum dots (CdS or PbS) as signal tags. PV is a dendrimer enzyme linked polymer, which can immobilize abundant QDs to amplify the stripping voltammetry signals from the metal ions (Pb or Cd). The capture probes were prepared through the immobilization of SH-labeled oligonucleotides, which can complementary with HIV and TB DNA, on the magnetic Fe 3 O 4 @Au (GMPs) beads. After sandwich-hybridization, the polymer nanotracers together with HIV and TB DNA targets were simultaneously introduced onto the surface of GMPs. Then the two encoding metal ions (Cd 2+ and Pb 2+ ) were used to differentiate two viruses DNA due to the different subsequent anodic stripping voltammetric peaks at −0.84 V (Cd) and −0.61 V (Pb). Because of the excellent signal amplification of the polymer nanotracers and the great specificity of DNA targets, this assay could detect targets DNA as low as 0.2 femtomolar and exhibited excellent selectivity with the dynamitic range from 0.5 fM to 500 pM. Those results demonstrated that this electrochemical coding assay has great potential in applications for screening more viruses DNA while changing the probes.
研究领域:
生物医学工程
·
分子诊断学
·
纳米技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2021
·
IF 7.46
Wei Tang,Ruo Mi,Lun Wang,Hongqi Chen
A quantitative image analysis and separation system was designed for detection of human immunodeficiency virus (HIV) related DNAs in the red region by combining of upconversion luminescence nanoprobe and magnetic beads (MBs). Carboxyl-modified NaGdF 4 :Yb, [email protected] 4 upconversion nanoparticles (UCNPs) were connected to single-stranded DNA to form a reporter probe, while carboxyl-modified magnetic beads were connected to another single-stranded DNA to form a capture probe. In the presence of target HIV DNAs, NaGdF 4 :Yb, [email protected] 4 UCNPs and magnetic beads were conjugated to form UCNPs-DNA-MBs nanocomplexes through DNA-DNA heteroduplexes based on the principle of complementary base pairing. After magnetic separation, the upconversion luminescence intensity of UCNPs-DNA-MBs nanocomplexes was related to the concentration of the target HIV DNAs. The detection methods were investigated by fluorescence spectrometry and quantitative image analysis based on upconversion luminescence total internal reflection (TIR) imaging platform. Compared to the luminescence spectral analysis, the quantitative image analysis shows higher sensitivity (linear range of 0.02−30 nM and detection limit of 0.0102 nM). More importantly, this method only needs 10 μL sample and the analysis time is about 0.1 s. The method combines quantitative image analysis with magnetic separation technology, which has the characteristics of low background interference and high separation efficiency allowing for application to the field of biology.
研究领域:
分析化学
·
生物技术
·
纳米技术
·
病毒学
✓ 已验证
DOI
高影响因子
TALANTA
·
2020
·
IF 5.339
Jiao Zhou,Lei Han,Yu Ling,Lei Wang,Nian Bing Li,Hong Qun Luo
Relying on the specific coordination of Ag + and mismatched cytosine-cytosine (C–C), the high-efficiency inhibition of urease by Ag + ion, and the rapid and sensitive response of phenol red to pH, a sensitive ratiometric sensor has been designed for visual detection of human immunodeficiency virus gene (HIV DNA). This sensor utilizes the HIV DNA to initiate catalytic hairpin assembly (CHA) process, releasing Ag + to inhibit subsequent urease-catalyzed urea hydrolysis and prevent the pH of the solution from rising. The CHA process and the absorbance ratio of phenol red at different wavelengths (A 559 /A 432 ) amplify the signal, allowing the sensor to detect HIV DNA from 10 to 130 nM in a sensitive and highly selective manner with a low detection limit of 7.8 nM. In addition, this sensor can visually distinguish different concentrations of HIV DNA within a certain range and possesses a good recovery in 1% of serum samples, which will provide new ideas for biosensor design, dipstick test, blood test, and other clinical disease prevention.
研究领域:
生物技术
·
纳米技术
·
分析化学
·
分子生物学
·
医学诊断
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2018
·Vol. 411
·期 19
·4569-4576
·
IF 3.307
Wang Kun,Zhai Fu-Heng,He Meng-Qi,Wang Jin,Yu Yong-Liang,He Rong-Huan
A simple fluorescence biosensor is developed based on the enzyme-assisted cascade amplification strategy. The amplification system consists of a hairpin-structure DNA (H-DNA) and exonuclease III. The target DNA can hybridize with the H-DNA and initiate exonuclease III-assisted target recycling amplification to generate abundant G-rich DNA (G-DNA). One region of G-DNA is designed to possess the same sequence as target DNA. Thus, the G-DNA can also hybridize with H-DNA and initiate the digestion of H-DNA. The cascade strategy in this amplification system causes the concentration of G-DNA to grow exponentially. The fluorescence intensity of N -methylmesoporphyrin IX (NMM) is highly enhanced due to the formation of G-quadruplex configuration. Under optimal conditions, the cascade system could achieve an admirable sensitivity with a detection limit of 52 fM for HIV DNA, and guarantees a satisfactory specificity. Moreover, the cascade system could be implemented for other target DNA detections by substituting the recognition region of the H-DNA. In this way, a detection limit of 65 fM for HBV DNA could be achieved by the cascade system. The target DNA analysis in a real serum sample further indicates that this biosensor has potential for future application in clinical diagnosis.
A simple and label-free cascade amplification strategy is developed by exploiting hairpin DNA and EXO III for sensitive DNA detection.
研究领域:
生物化学
·
分子生物学
·
分析化学
·
临床诊断
✓ 已验证
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）