TALANTA
·
2020
·
IF 5.339
Xiaoyan Feng,Xiangyu Meng,Fangbin Xiao,Zoraida P. Aguilar,Hengyi Xu
Sepsis caused by bacteria has high morbidity and mortality, and it is neccerssay to establish a fast, convenient, and facility assays for detection of bacteria. In this study, we have developed established a simple, rapid, and ultrasensitive vancomycin (Van) and dendrimer nanoparticles-based method to isolate and detect bacteria in human blood using a multivalent binding strategy. The proposed Bio-den-Van multivalent capture nanoplatform combined with m-qPCR for simultaneous detection of two kinds of bacteria was demonstrated with rapid 2 min bacteria isolation with a linear range at 3.2 × 10 1 –3.2 × 10 6 CFU·mL -1 for L. monocytogenes and 4.1 × 10 1 –4.1 × 10 6 CFU·mL -1 for S. aureus , respectively. The limit of detection (LOD) for simultaneous detection of L. monocytogenes and S. aureus were 32 and 41 CFU·mL -1 in spiked human blood samples, respectively. Other bacteria had an insignificant interference with the test results. This Bio-den-Van multivalent capture nanoplatform combined with m-qPCR detection exhibited rapid, high sensitivity and specificity in simultaneous detection of various bacteria. To our knowledge, this is the first time that Bio-den-Van multivalent capture nanoplatform was used with Van as a recognition molecule for the simultaneous capture and subsequent detection of two bacteria from spiked human blood sample. This method holds great potential for future clinical applications.
研究领域:
医学诊断
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纳米技术
·
分子生物学
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临床微生物学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2019
·
IF 6.35
Kun Yin,Xi Zeng,Weizhi Liu,Yakun Xue,Xingrui Li,Wei Wang,Yanling Song,Zhi Zhu,Chaoyong Yang
As the third-generation nucleic acid amplification technology, digital polymerase chain reaction (PCR) has been widely adopted in the analysis of nucleic acids. However, further application of this powerful technology is hindered due to the limitation of surfactants. Here, for the first time, we propose the use of colloidosomes self-assembled from fluorinated silica nanoparticle for digital PCR to address this limitation. A one-step fluorinated silica nanoparticle synthesis method is proposed, which is much more convenient and reproducible compared with the synthesis of conventional fluorine-based surfactants. Fluorinated silica nanoparticles facilitate the formation of colloidosomes with excellent stability capable of enduring the rapid temperature changes associated with the PCR and avoiding material exchange (cross-talk) between droplets for high-fidelity analysis. The colloidosome digital PCR method was developed using these colloidosomes as highly parallel reactors for single-copy nucleic acid amplification and rare mutant detection. The method is robust and accurate, and it offers possibilities for a great variety of applications, such as gene expression studies, single cell analysis, and circulating tumor DNA detection.
研究领域:
化学
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纳米技术
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生物技术
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分子生物学
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医学诊断
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产品关联
DOI
PubMed
高影响因子
COLLOIDS AND SURFACES A-PHYSICOCHEMICAL AND ENGINEERING ASPECTS
·
2022
·
IF 5.518
Yanan Jia,Lin Hao,Jun Zhang,Xing Zhang,Siying An,Ming Su,Huan Wang,Yufan Zhang
We describe a PCR-free, enzyme-free, and simple biosensing platform for the electrochemical monitoring of telomerase activity based on Au nanorods (AuNRs) as an electrocatalytic redox mediator, which enhances the electrochemical signal by catalyzing acetaminophen (AP). In this process, acid-functionalized ordered mesoporous carbon (OMC) was modified on a glassy carbon electrode (GCE) surface to offer -COOH sites. Moreover, the amino-modified S1 was fixed on the modified electrode by conducting an amide reaction. TS is extended to (TTAGGG) n repeat units in the presence of telomerase extract and dNTPs. One end (5′) is hybridized with S1 on the electrode, and the remaining end (3′) is combined with AuNRs-S2 probe. Clearly, the telomerase activity is relevant to the amount of connected AuNRs, resulting in different response signals. Therefore, with the telomerase extension triggered hybridization reaction, a large amount of AuNRs catalyst was introduced onto the surface of GCE as the sensing platform, thus achieving a sensitive electrochemical telomerase activity assay in the range of 20–2.15 × 10 6 HeLa cells mL −1 . This sensing platform shows that the limit of detection is 1.74 HeLa cells mL −1 , thus providing a promising method for the determination of telomerase activity and a powerful biosensor for accurate cancer monitoring and diagnosis.
研究领域:
生物传感器
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电化学
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癌症诊断
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纳米技术
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产品关联
DOI
高影响因子
MICROCHIMICA ACTA
·
2018
·Vol. 185
·期 8
·1-7
·
IF 5.705
Meng Fanyu,Xu Yuanyuan,Dong Wenfei,Tang Yuguo,Miao Peng
The paper describes a voltammetric method for the quantitation of the activity of telomerase extracted from cancer cells. A thiolated single-stranded telomerase substrate primer was firstly immobilized on a gold electrode. In the presence of a mixture of telomerase and deoxynucleotide triphosphates, the primer becomes elongated and contains repetitive nucleotide sequences (TTAGGG) n . After hybridization with blocker DNA, gold nanoparticles are added and captured by the elongated single-stranded DNA. This reduces the charge transfer resistance of the gold electrode. The telomerase activity is then quantified via differential pulse voltammetry, typically at 0.12 V (vs. SCE). The method is PCR-free, rapid, and convenient. It was applied to the detection of HeLa cells via the telomerase activity of lysed cells. The detection range was from 500 to 50,000 cells/mL and the detection limit was as low as 500 cells/mL.
A telomerase substrate (TS) primer is immobilized on a gold electrode as the sensing interface to detect the activity of telomerase extracted from cancer cells. Unmodified gold nanoparticles (AuNPs) are utilized which change the electrochemical responses.
研究领域:
生物化学
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分析化学
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癌症研究
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DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2025
·
IF 7.7
Jiahui Li,Yuanyuan Zhang,Zhengshun Wen,Jiangbing Shuai,Xiaofeng Zhang,Jinru Zhou,Aiqian Ye,Linglin Fu
The delayed reporting of milk allergen residue detection results is a significant factor contributing to substantial losses for food production enterprises. Cumbersome operational procedures and high testing costs often lead to the abandonment of tests, resulting in frequent allergic reactions among milk-sensitive populations. Therefore, there is an urgent need for a novel, rapid, and low-cost detection method to mitigate enterprise losses and protect allergic individuals. This study investigates the formation mechanism of convection PCR, driven by Fast Taq polymerase in standard PCR tubes, and develops a fluorescence detection device for convection PCR. Through molecular docking and experimental validation of the trimmed β-LG aptamers, we elucidate their potential multi-site binding mechanism. Utilizing this mechanism, we designed a dual-strand hybridization magnetic bead detection system (dsA-SA-B), employing easily synthesized and amplified aptamer complementary strands as an indicator signal; Signal amplification was achieved within 15 min, while the entire detection process was completed in less than 50 min. This detection method allows for the rational selection of aptamer sequences based on actual testing needs, enabling the detection of hydrolyzed milk powder. The method demonstrated a linear detection range for β-LG from 5 to 100 μg/mL, with a detection limit as low as 0.25 μg/mL, effectively identifying hydrolyzed milk powder. This approach transforms protein detection into nucleic acid detection, and lays a solid foundation for multiplex PCR detection of eight major food allergens, facilitating the widespread detection of milk allergens and ultimately contributing to improved food safety and public health.
研究领域:
分子生物学
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食品安全
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免疫学
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生物技术
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分析化学
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DOI
高影响因子
FOOD CONTROL
·
2022
·
IF 6.652
Xiaoran Li,Ruohong Zhang,Chun Wang,Xiaofang Wang,Yang Yang,Shenghui Cui,Yunchang Guo
Listeriosis outbreaks caused by contaminated leafy greens leaf have increased in recent years, which can pose a serious threat to public health. Thus, a rapid, precise, and effective detection method is urgently needed. We reported a novel sample pre-treatment that utilizes β-cyclodextrin and milk protein-coated activated charcoal to remove PCR inhibitors in leafy green matrices and facilitate the recovery of L. monocytogenes , which can be coupled with PCR subsequently. The primers targeting the iap gene of L. monocytogenes were used for testing the specificity with 141 bacterial strains. Artificially contaminated leafy greens (10 0 –10 5 CFU/25 g) that include spinach, lettuce, Lactuca sativa L., cole, and cabbage were tested to determine the sensitivity of the developed method. Natural leafy greens were tested to evaluate the application of the developed method compared with the conventional culture method. The stability (shelf life) of the reagents used in the developed method was determined using reagents stored at −20 °C for 3, 6, and 12 months to evaluate the sensitivity and application of the method. As a result, the specificity of the developed method was 100% and the detection limit was 10 1 CFU/25 g. The total detection time without pre-enrichment was 4 h. The detection rate of L. monocytogenes (spinach 5%, lettuce 3%, cole 1%, Lactuca sativa L. 0%, and cabbage 0%) and the detection rate of viable bacteria (100%) were consistent in both diagnostic methods (the coincidence rate was 100%). Furthermore, the sensitivity and the application of the developed method were not influenced using the reagents stored for 3, 6, and 12 months. The reagents could be used normally after storing at −20 °C for at least 12 months, which proved that these reagents have good stability under freezing conditions. This method for the detection of L. monocytogenes was rapid, sensitive, and specific, and can potentially be used to detect and monitor the target pathogen in leafy greens and to reduce the potential risk of L. monocytogenes for public health.
研究领域:
食品安全
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微生物学
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DOI
高影响因子
ANALYTICAL BIOCHEMISTRY
·
2020
·
IF 2.877
Lijun Wang,Tianyu Liao,Hong Zhou,Yukun Huang,Pengfei Chen,Xiao Yang,Xianggui Chen
Development of a rapid and sensitive method for Salmonella spp. detection is of great importance for ensuring food product safety due to its low infective dose. In this study, a colorimetric method based on the peroxidase-like activity of Cu(II)-modified reduced graphene oxide nanoparticles (Cu 2+ -rGO NPs) and PCR was successfully developed to detect Salmonella spp. in milk. Under optimal conditions, the developed colorimetric method exhibited high sensitivity and strong specificity for Salmonella spp . detection. The limit of detection was 0.51 CFU/mL with a linear range from 1.93 × 10 1 to 1.93 × 10 5 CFU/mL. A specificity study demonstrated that this method can specifically distinguish Salmonella typhimurium and Salmonella enteritidis from other foodborne pathogens. The application of the proposed method for milk sample detection was also validated, and the recovery rates of S . typhimurium in spiked milk sample ranged from 102.84% to 112.25%. This colorimetric sensor exhibits enormous potential for highly sensitive detection of bacteria in milk sample.
研究领域:
食品科学
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纳米技术
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微生物学
·
分析化学
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产品关联
DOI
PubMed
高影响因子
Practical Laboratory Medicine
·
2023
·
IF 1.9
Xu Tang,Lin-Ping Fan,Yang Liu
Introduction Group B streptococcus(GBS)often causes adverse outcomes such as urinary system infection, intrauterine infection, premature birth, and stillbirth in perinatal women. Perinatal screening of GBS is conducive to guiding clinical scientific intervention and improving delivery outcomes.This study quantitative real-time PCR (RT-qPCR) combined with magnetic separation was used for GBS detection. Materials and methods Sample pre-treatment in this study involved the utilization of magnetic separation (MS) technology, aiming to expedite the detection process and enhance detection sensitivity, and the cfb gene of group B streptococcus was used as the target gene to establish quantitative real-time PCR (RT-qPCR) to detect group B streptococcus. Results It was found that penicillin-functionalized magnetic beads had a good ability to enrich and capture group B Streptococcus.The findings revealed an exceptional detection sensitivity, with the ability to detect B streptococcus in urine samples at levels as low as 10 2 CFU/mL. Conclusions The utilization of MS technology in conjunction with the RT-qPCR (MS-RT-qPCR) assay, as demonstrated in this study, offers a viable approach for prenatal screening of group B streptococcus among perinatal women.
研究领域:
微生物学
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分子生物学
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妇产科学
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临床诊断
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DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2018
·Vol. 185
·期 9
·1-9
·
IF 5.705
Wang Jing,Li Haigang,Li Tingting,Ling Liansheng
A one-step, one-tube colorimetric assay is described for the detection of bacterial double-stranded DNA (dsDNA). It utilizes a G-quadruplex DNAzyme produced by an asymmetric polymerase chain reaction (As-PCR) that catalyzes the oxidation of cysteine to form cystine. This results in the formation of oligonucleotide-modified gold nanoparticles via triplex formation, and eventually in a color change from red to blue that occurs within about 10 mins. This can be measured by ratiometric colorimetric (at 525 and 600 nm). The limit of detection (LOD) for the model analyte (dsDNA of Staphylococcus aureus ( S. aureus )) is as low as 0.28 pg per 0.05 mL with a good linear response ranging from 16.0 fg·μL −1 to 1.6 ng·μL −1 . This is much lower than previously reported LODs. The assay is highly selective for S. aureus dsDNA over a range of other bacterial DNAs. Conceivably, it provides an attractive alternative tool for rapid detection of bacterial dsDNA as required in pathogen screening in the food industry.
Schematic presentation of a colorimetric assay for bacterial DNA. It is based on the catalytic activity of a G-quadruplex DNAzyme that is formed by an asymmetric PCR involving triplex DNA formation and gold nanoparticle (AuNPs) aggregation.
研究领域:
分子生物学
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纳米生物技术
·
病原体检测
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2020
·Vol. 412
·期 17
·4155-4163
·
IF 3.637
Li Jinge,Zhao Liying,Wen Dongxiao,Li Xiaofei,Yang Huaixia,Wang Dazhong,Kong Jinming
In this work, a new method of CYFRA21-1 DNA (tDNA) detection based on electrochemically mediated atom transfer radical polymerization (e-ATRP) and surface-initiated reversible addition-fragmentation chain transfer polymerization (SI-RAFT) cascade polymerization and AgNP deposition is proposed. Firstly, the peptide nucleic acid (PNA) probe is captured on a gold electrode by Au-S bonds for specific recognition of tDNA. After hybridization, PNA/DNA strands provide high-density phosphate groups for the subsequent ATRP initiator by the identified carboxylate-Zr 4+ -phosphate chemistry. Then, a large number of monomers are successfully grafted from the DNA through the e-ATRP reaction. After that, the chain transfer agent of SI-RAFT and methacrylic acid (MAA) are connected by recognized carboxylate-Zr 4+ -carboxylate chemistry. Subsequently, through SI-RAFT, the resulting polymer introduces numerous aldehyde groups, which could deposit many AgNPs on tDNA through silver mirror reaction, causing significant amplification of the electrochemical signal. Under optimal conditions, this designed method exhibits a low detection limit of 0.487 aM. Moreover, the method enables us to detect DNA at the level of PCR-like and shows high selectivity and strong anti-interference ability in the presence of serum. It suggests that this new sensing signal amplification technology exhibits excellent potential of application in the early diagnosis of non-small cell lung cancer (NSCLC).
Electrochemical detection principle for CYFRA21-1 DNA based on e-ATRP and SI-RAFT signal amplification technology.
研究领域:
生物化学
·
分析化学
·
癌症诊断
·
纳米技术
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产品关联
DOI
PubMed
高影响因子
LAB ON A CHIP
·
2023
·
IF 6.1
Yulin Ren,Jingcheng Ji,Haoqing Zhang,Lei Cao,Jie Hu,Feng Xu,Zedong Li
Droplet digital polymerase chain reaction (ddPCR) technology has found widespread applications in the ultrasensitive analysis of nucleic acids, where integrated ddPCR platforms with the capability of sample dispersion, followed by in situ amplification and data analysis, are highly expected. However, current integrated ddPCR platforms are usually limited by either difficultly mass-produced materials or lack of integrated control instruments, restricting their practical application. This paper proposes an integrated three-in-one ddPCR platform with high user-friendliness and practicability, which is composed of an easy-to-use chip and a matching control instrument. The chip was made of thermally resistant and easily mass-produced polycarbonate (PC) material, and the benchtop control instrument was designed to perform droplet generation, in situ amplification, and fluorescence reading. The droplet generation and in situ heating on the chip were well characterized. Finally, the performance of the platform was validated through the analysis of the EGFR L858R mutation in lung cancer. The proposed three-in-one ddPCR platform shows great practicability in ultrasensitive nucleic acid testing. By virtue of its sensitivity, practicability, and cost-effectiveness, the ddPCR can serve as a universal detection platform for monitoring nucleic acid in the fields of tumor diagnosis, pathogen detection, and prenatal diagnosis.
研究领域:
生物技术
·
分子生物学
·
医学诊断
·
基因工程
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产品关联
DOI
PubMed
高影响因子
LAB ON A CHIP
·
2019
·Vol. 19
·期 16
·2663-2668
·
IF 6.914
Zhenqing Li,Ruixue Ju,Shinichi Sekine,Dawei Zhang,Songlin Zhuang,Yoshinori Yamaguchi
Current continuous flow polymerase chain reaction (CF-PCR) microfluidic chips require external precision syringe pumps and off-line methods (e.g., electrophoresis and hybridization) to detect PCR products, resulting in complex operations and possible cross-contamination and consequently CF-PCR is still confined to laboratories. Herein, a portable all-in-one microfluidic device is fabricated for rapid diagnosis of pathogens based on an integrated CF-PCR and electrophoresis biochip. A new method was proposed for automatic sample injection into the chip which can substitute the costly external precision syringe pump. It not only achieves rapid DNA amplification and on-site PCR product detection, but also realizes automatic sample injection. As an application, three periodontal pathogens (e.g., Porphyromonas gingivalis, Treponema denticola and Tannerela forsythia) were successfully amplified in the device. Treponema denticola was amplified in as short as 2′31′′, and detection of PCR products was completed within 3′43′′. The minimum number of bacteria that can be amplified was 125 cfu per μl. The all-in-one device has the potential to be applied in point-of-care nucleic acid testing for diseases.
研究领域:
微流控技术
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生物医学工程
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分子诊断
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即时检测
✓ 已验证
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）