Frontiers in Microbiology
·
2018
·
IF 4.019
Yu Guanliu,Yu Xianglong,Yang Guoping,Tang Yi,Diao Youxiang
Duck Tembusu virus (DTMUV) is an emerging pathogenic flavivirus that has resulted in large economic losses to the duck-rearing industry in China since 2010. Therefore, an effective diagnostic approach to monitor the spread of DTMUV is necessary. Here, a novel diagnostic immunochromatographic strip (ICS) assay was developed to detect DTMUV. The assay was carried out using colloidal gold coated with purified monoclonal antibody A12D3 against envelope E protein. Purified polyclonal C12D1 antibodies from BALB/c mice against the envelope E protein were used as the capture antibody. Goat anti-mouse IgG was used to detect DTMUV, which was also assembled on the ICS. Results showed that the ICS could specifically detect DTMUV within 10 min. It also could be stored 25 and 4°C for 4 and 6 months, respectively. The sensitivity of the ICS indicated that the dilution multiples of positive allantoic fluid of DTMUV (LD50: 104.33/0.2 ml) was up to 200. Its specificity and sensibility showed no significant change under the above storage situations. Fifty clinical samples were simultaneously detected by ICS and reverse-transcription polymerase chain reaction with a 93.9% coincidence rate between them. It proved that the ICS in the present study was highly specific, sensitive, repeatable, and more convenient to rapidly detect DTMUV in clinical samples.
研究领域:
兽医学
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病毒学
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诊断技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2024
·Vol. 191
·期 4
·1-10
·
IF 5.3
Jin Birui,Ma Chuan,Zhang Chuyao,Yin Huiling,Zhao Guoxu,Hu Jie,Li Zedong
There is an urgent need for a point-of-care testing (POCT) method in developing and underserved regions to distinguish between two Monkeypox virus (MPXV) clades, given their varying transmissibility and clinical manifestations. In this paper, we target the specific complement protein gene fragment of two MPXV clades and construct a high-performance upconversion nanoparticles–based lateral flow assay (UCNPs-based LFA) with double T-lines and a shared C-line. This enables qualitative and quantitative dual-mode detection when combined with a smartphone and a benchtop fluorescence analyzer. The developed LFA exhibits stable performance, convenient operation, rapid readout (within 8 min), and a much lower limit of detection (LOD) (~ pM level) compared to existing POCT methods. The proposed detection platform demonstrates significant potential for pathogen diagnosis using a POCT approach. Graphical Abstract
研究领域:
病毒学
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诊断方法
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纳米技术
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即时检测
✓ 已验证
产品关联
DOI
PubMed
高影响因子
LANGMUIR
·
2024
·
IF 3.7
Yaqin Zhang,Jianai Chen,Fange Kong,Chunxia Wang,Hangyu Guo,Yingchun Li,Jiahui Lu,Jicheng Zhang,Jiasi Wang,Yulin Zhou
Viral infections, such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), are some of the most dangerous threats to humans. SARS-CoV-2 has caused a global pandemic, highlighting the unprecedented demand for rapid and portable diagnostic methods. To meet these requirements, we designed a label-free colorimetric platform that combines the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated proteins (Cas) 12a system for naked-eye detection (named LFP). This method utilizes reverse transcription loop-mediated isothermal amplification (RT-LAMP) and the trans-cleavage activity of the CRISPR/Cas12a system to increase the sensitivity and specificity of the reaction. This platform can detect as few as 4 copies/μL of RNA and produces no false positive results when tested against the influenza virus. To better meet the requirements of point-of-care (POC) detection, we developed a portable device that can be applied in resource-poor and densely populated regions. The LFP assay holds great potential for application in resource-limited settings, and the label-free gold nanoparticle (AuNPs) probe can reduce costs, making it suitable for large-scale screening. We expect that the LFP assay will be promising for the POC screening of COVID-19.
研究领域:
分子生物学
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生物技术
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病毒学
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纳米技术
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诊断方法
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICAL BIOCHEMISTRY
·
2024
·
IF 2.6
Yaling Yuan,Yue Zhong,Yanbin Yang,Ke Li
Myoglobin (Myo), creatine kinase-MB (CKMB), and cardiac troponin I (cTnI) are crucial biomarkers for diagnosing acute myocardial infarction (AMI) The accurate and rapid detection of these three targets can greatly improve the prognosis of AMI patients. Herein, this study developed a microfluidic immunofluorescence method that can detect all three targets in 10–15 min. Ultrasonic atomization and spray technology are used to modify the surface of the injection-molded microfluidic chip (MFC), which effectively solves the problem of biological cross-linking and antibody immobilization on the MFC surface. In addition, it improves the hydrophilicity of the chip surface, thus enhancing fluid self-driving effect. The linear response towards Myo, CKMB and cTnI range from 5 ng/mL to 500 ng/mL, 1 ng/mL to 70 ng/mL, and 0.05 ng/mL to 30 ng/mL, respectively. The intra-batch precision is ≤ 10%, and the inter-batch precision is ≤ 15%. Furthermore, this method shows good consistency compared with the BECKMAN ACCESS2 chemiluminescent immunoanalyzer. The present work provides an AMI diagnostic method with high sensitivity, good repeatability, high accuracy and simple operation, which can satisfy the needs of clinical diagnosis, and shows promising application prospects.
研究领域:
生物医学工程
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心脏病学
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诊断方法
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微流控技术
✓ 已验证
DOI
PubMed
高影响因子
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
·
2024
·
IF 3.1
Yan Na,Hu Zhiwei,Zhang Lingling
The development of a bio-sensing strategy based on CRISPR/Cas that is exceptionally sensitive is crucial for the identification of trace molecules. Colorimetric miRNA detection utilizing CRISPR/Cas13a-triggered DNAzyme signal amplification was described in this article. The developed strategy was implemented for miRNA-21 detection as a proof of concept. The cleavage activity of Cas13a was triggered when the target molecule bonded to the Cas13a-crRNA complex and cleaved uracil ribonucleotides (rU) in the substrate probe. As a consequence, the S chain was liberated from the T chain that had been modified on magnetic beads (MB). The G-rich sections were then exposed when the catalytic hairpin assembly between the H1 and H2 probes was activated by the released T@MB. G-rich section can fold into G-quadruplex. By catalyzing the formation of green ABTS 3 – via HRP-mimicking G-quadruplex/hemin complexes, colorimetric measurements of miRNA can be achieved visually through DNAzyme-mediated signal amplification. The method demonstrated a low limit of detection of 27 fM and a high selectivity towards target miRNA eventually. As a result, the developed strategy provides a clinical application platform for the detection of miRNAs that is both ultrasensitive and extremely specific.
研究领域:
生物技术
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分子生物学
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诊断方法
✓ 已验证
DOI
PubMed
高影响因子
Frontiers in Bioengineering and Biotechnology
·
2025
·
IF 4.8
Xiao Xu,Anji Zhu,Lixia Bai,Shihong Li,Wenbo Ge,Yajun Yang,Xiwang Liu,Zhe Qin,Zhun Li,Jianyong Li
Introduction Gram-negative bacteria are key pathogens causing food contamination and animal diseases (such as bovine mastitis), and they are prone to developing multidrug resistance, necessitating rapid and sensitive detection technologies. Methods We developed a novel detection method based on an antibiotic affinity strategy, which combines vancomycin-functionalized magnetic beads (Van-MBs) with aminopeptidase test strips. Vancomycin, which binds exclusively to D-alanyl-D-alanine (D-Ala-D-Ala) on the cell wall of Gram-positive bacteria, is used by Van-MBs to capture Gram-positive bacteria from the sample. After magnetic separation, the supernatant containing Gram-negative bacteria is retained. This supernatant reacts with aminopeptidase test strips, utilizing the enzyme's unique properties as a Gram-negative enzyme. Visual detection is made possible by the enzyme's catalysis of the chromogenic substrate, which produces a yellow signal. Results Using raw milk as a representative sample, validation showed that the approach achieves a detection limit of 1.0 × 10 1 CFU/mL within 100 minutes. Discussion This approach is quick, sensitive, and visual, and it doesn't require complicated equipment. It also offers useful technical support for safe food production, early animal disease detection, and sensible antibiotic administration.
研究领域:
微生物学
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食品安全
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诊断方法
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传染病
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抗菌素耐药性
✓ 已验证
产品关联
DOI
PubMed
高影响因子
CURRENT PHARMACEUTICAL BIOTECHNOLOGY
·
2015
·Vol. 16
·期 8
·716-723
·
IF 1.95
Hui Yu,Guangying Zhao,Wenchao Dou
Aimed to explore an agglutination test which can simultaneously detect two pathogenic bacteria, an agglutination test based on colored silica nanoparticles (colored-SiNps) was established in this work. Monodisperse colored-SiNps were used as agglutination test carriers; red-SiNps and blue-SiNps were prepared by reverse microemulsion with C.I. Reactive red 136 and C.I. Reactive Blue 14. Then the red-SiNps were sensitized with antibodies against E. sakazaki and denoted as IgG-red-SiNps; The blue-SiNps were coated with antibodies against S. pullorum and S. Gallinarum and denoted as IgGblue- SiNps. The mixture solution of IgG-red-SiNps and IgG-blue-SiNps could simultaneously agglutinate with E. sakazakii and S. pullorum and S. gallinarum on glass slide. The E. sakazakii and S. pullorum and S. gallinarum could be simultaneously detected by agglutination test with obvious agglutination phenomena. The E. sakazakii and S. pullorum and S. gallinarum could both be detected in a range from 4×103 to 4×109 CFU/mL. The pullorum and S. gallinarum and E. sakazakii in the infected food sample were detected by mixture solution of IgG-red-SiNps and IgG-blue-SiNps too. This agglutination test was easy and rapid, it might be useful for in situ rapid detection method for simultaneously screening different pathogenic microorganisms of foods and feeds in the field.
研究领域:
食品安全
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纳米生物技术
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微生物检测
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诊断方法
✓ 已验证
DOI
高影响因子
Frontiers in Cellular and Infection Microbiology
·
2025
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IF 4.8
Yihui Xie,Zhaoqin Zhu,Yuxin Shi,Zihui Zhao,Xueqin Qian,Yinzhong Shen
Objective The BACTEC MGIT 960 system is currently the most widely used rapid culture system for Mycobacterium tuberculosis . However, its reagents have complex compositions, expensive, and rely on import. This study aimed to develop a self-formulated culture with simple composition, low cost, and performance comparable to or even better than that of the BACTEC MGIT 960 system. Methods Five experimental media were formulated by supplementing Sauton medium with carrot extract, potato extract, oleic acid, or choline chloride. These were compared against commercial MGIT 960 medium using M. tuberculosis H37Rv suspensions (10 - ³-10 -6 mg/mL, triplicate cultures). Time to detection (TTD) and growth index (GI) were monitored automatically using the BACTEC MGIT 960 system. Results For medium-high inocula (10 - ³-10 -5 mg/mL), carrot- and potato-supplemented Sauton media showed equivalent performance to MGIT 960 (1-day longer TTD, P>0.05; comparable GI values). At low inoculum (10 -6 mg/mL), MGIT 960 failed to detect growth within 42 days, whereas supplemented Sauton medium achieved positivity within 30 days. The carrot extract medium performed best (mean TTD = 20.2 days), followed by oleic acid and oleic acid + choline chloride formulations. Conclusion Carrot- and potato-supplemented Sauton medium demonstrated comparable performance to the MGIT 960 system for specimens with medium-to-high bacterial loads. Carrot extract, oleic acid, and choline chloride significantly enhanced the detection rate of low-inoculum, with carrot extract exhibiting the most pronounced growth-promoting effects. This study validates an accessible alternative to expensive imported media, particularly beneficial for resource-limited settings.
研究领域:
微生物学
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临床检验科学
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结核病研究
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诊断方法
✓ 已验证
产品关联
DOI
PubMed
高影响因子
METHODS
·
2021
·
IF 3.608
Menglu Hu,Debin Zhu,Xiaoming Zhou
The construction of a rapid, simple, and specific nucleic acid detection platform is of great significance to the control of the large-scale spread of infectious diseases. We have recently established a magnetic pull-down-assisted colorimetric method based on the CRISPR/Cas12a system (termed M−CDC), which effectively integrates the advantages of CRISPR/Cas12a, magnetic beads-based separation, and AuNP bioprobe to provide a simple and specific biosensing platform for nucleic acid assay. The M−CDC method is compatible with point-of-care testing and enables the detection of nucleic acid samples in less than an hour without relying on expensive and complex instruments. In this paper, step-by-step instructions for M−CDC assay, including recombinase polymerase amplification (RPA)/reverse transcription-polymerase chain reaction (RT-RPA) of DNA or RNA, Cas12a-mediated target recognition and cleavage, and subsequent magnetic beads-mediated colorimetric readouts are provided. In addition, the protocol for the expression and purification of Lachnospiraceae bacterium -Cas12a (LbCas12a) protein, the design and synthesis of high-efficient crRNA, and the preparation of AuNP bioprobe are also offered.
研究领域:
生物技术
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分子生物学
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诊断方法
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纳米技术
✓ 已验证
DOI
PubMed
高影响因子
MATERIALS & DESIGN
·
2022
·
IF 9.417
Lihua Li,Menglin Song,Xinyue Lao,Sin-Yi Pang,Yuan Liu,Man-Chung Wong,Yingjin Ma,Mo Yang,Jianhua Hao
Here, we firstly introduce a detection system consisting of upconversion nanoparticles (UCNPs) and Au nanorods (AuNRs) for an ultrasensitive, rapid, quantitative and on-site detection of SARS-CoV-2 spike (S) protein based on Förster resonance energy transfer (FRET) effect. Briefly, the UCNPs capture the S protein of lysed SARS-CoV-2 in the swabs and subsequently they are bound with the anti-S antibodies modified AuNRs, resulting in significant nonradiative transitions from UCNPs (donors) to AuNRs (acceptors) at 480 nm and 800 nm, respectively. Notably, the specific recognition and quantitation of S protein can be realized in minutes at 800 nm because of the low autofluorescence and high Yb-Tm energy transfer in upconversion process. Inspiringly, the limit of detection (LOD) of the S protein can reach down to 1.06 fg mL −1 , while the recognition of nucleocapsid protein is also comparable with a commercial test kit in a shorter time (only 5 min). The established strategy is technically superior to those reported point-of-care biosensors in terms of detection time, cost, and sensitivity, which paves a new avenue for future on-site rapid viral screening and point-of-care diagnostics.
研究领域:
生物医学工程
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纳米技术
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病毒学
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即时检测
·
诊断方法
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2019
·
IF 6.35
Qiuyuan Lin,Xin Ye,Zhipeng Huang,Bin Yang,Xueen Fang,Hui Chen,Jilie Kong
Cyclooxygenase-2 (COX2) mRNA represents a key biomarker for identifying subjects with colorectal cancer (CRC), while there is still no rapid and sensitive detection method for COX2 mRNA. Loop-mediated isothermal amplification (LAMP) is extensively developed for the amplification of nucleic acids; however, its application is frequently hindered by serious nonspecific amplification. Herein, this work reported a graphene oxide (GO)-based LAMP method to enable the one-step detection of COX2 mRNA in cancer cells and serum samples. We found that GO greatly enhanced the specificity of LAMP through decreasing nonspecific hybridization and the fluorescence background signal because of the simultaneous adsorption of single-stranded primers and DNA staining dyes on GO. The detection limit of developed GO-based LAMP was 2 orders of magnitude more sensitive compared to that of classical LAMP. Then a GO-based reverse transcription (RT)-LAMP strategy was further developed and applied to detect COX2 mRNA in CRC cancer cells and serum samples with high specificity. The GO-based LAMP platform with advantages of low cost, simplicity, high specificity, and sensitivity holds considerable potential for real-time fluorescence monitoring of nucleic acid amplification in a wide range of fields.
研究领域:
分子生物学
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癌症研究
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生物技术
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纳米技术
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诊断方法
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2022
·
IF 8.008
Yu Wang,Yu Xue,Qilin Zhao,Shuang Wang,Jian Sun,Xiurong Yang
Alzheimer’s disease (AD) is a common chronic neurodegenerative disease that manifests as cognitive impairment and behavioral deficits and severely threatens the health of the elderly. Acetylcholinesterase (AChE) plays a vital role in biological signaling and is an essential target for the early diagnosis and treatment of AD. Herein, 2D Zn-TCPP(Fe) nanosheets (NSs) employing Zn2+ and Fe-bound tetrakis(4-carboxyphenyl)porphyrin ligands were prepared through a surfactant-assisted synthetic method. The ultrathin two-dimensional (2D) metal–organic framework structures exhibited high peroxidase-like activity, which allowed the catalysis of the H2O2-initiated oxidation of colorless 3,3′,5,5′-tetramethylbenzidine (TMB) to blue oxidized TMB (ox-TMB). Such catalytic performance inspired us to develop a convenient, rapid, and sensitive acetylcholinesterase activity assay, during which AChE can catalyze the substrate acetylthiocholine (ATCh) to produce thiocholine (TCh), and TCh could especially enable the degradation of 2D Zn-TCPP(Fe) NSs accompanied by the reduction of ox-TMB production. Our proposed sensing system exhibited favorable selectivity and sensitivity (LOD of 0.029 mU/mL) and has excellent potential to evaluate AChE activity in human serum samples and to screen AChE inhibitors. This colorimetric assay could provide an alternative pathway for early diagnosis and drug screening of AD, facilitating the development of AD therapy.
研究领域:
神经退行性疾病
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生物化学
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纳米技术
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诊断方法
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药物筛选
✓ 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）