Analytical Methods
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2024
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IF 2.7
Huiqing Wang,Yang Li,Lin Tian,Xinyi Li,Qian Gao,Yaru Liu,Cuiping Ma,Qing Wang,Chao Shi
A concise and rapid detection method for Mycoplasma pneumoniae is urgently required due to its severe impact on human health. To meet such a need, this study proposed and constructed an innovative point-of-care testing (POCT) platform that consists of a hydrogen ion-selective loop-mediated isothermal amplification (H+-LAMP) sensor and an electrochemical detection device. The H+-LAMP sensor successfully integrated the working and reference electrodes and converted the H+ generated during the LAMP process into an electrochemical signal. High sensitivity and stability for pathogen detection were also achieved by treating the working electrode with an electrodeposited polyaniline solid contact layer and by using an ion-selective membrane. As a result, the sensor shows a sensitivity of 68.26 mV per pH, a response time of less than 2 s, and a potential drift of less than 5 mV within one hour, which well meets the urgent need. The results also demonstrated that the detection limit for Mycoplasma pneumoniae was lowered to 1 copy per μL, the nucleic acid extraction and detection process could be completed in 30 minutes, and the impact of interfering ions on the sensor was negligible. Validation with 20 clinical samples yielded satisfactory results. More importantly, the storage lifespan of such an electrochemical sensor is over seven days, which is a great advantage for on-site pathogen detection. Therefore, the hydrogen ion-selective sensor constructed in this investigation is particularly suitable as a core component for instant pathogen detection platforms.
研究领域:
生物医学工程
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微生物学
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电化学
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即时诊断
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DOI
PubMed
高影响因子
JOURNAL OF ECONOMIC ENTOMOLOGY
·
2025
·
IF 2.4
Cao Li-Xia,Pan Xing-Run,Zhang Wan-Na,Lu Jing-Bo,Hu Zhi-Kang,Sheng Cheng-Wang,Peng Ying-Chuan
The fall armyworm (Spodoptera frugiperda), a lepidopteran pest of highly destructive, has developed resistance to organophosphate and carbamate insecticides through target-site mutations, including A201S and F290V in the ace-1 gene encoding acetylcholinesterase (AChE). Developing rapid and reliable methods to detect these mutations is crucial for monitoring resistance and guiding effective management strategies. Although S. frugiperda populations remain sensitive to the tested insecticides, the A201S and F290V mutations were identified in collected field samples. To facilitate detection, a sensitive and field-deployable loop-mediated isothermal amplification (LAMP) assay was developed, targeting these mutations within the ace-1 gene of S. frugiperda. The specificity and sensitivity of the LAMP assay was optimized for rapid detection, with reactions performed at 68 °C for 80 min. Results were visualized through color change and confirmed by gel electrophoresis. Validation using DNA from laboratory and field samples demonstrated complete agreement between the LAMP assay and DNA sequencing, affirming the accuracy and reliability of the method. The developed LAMP assay provides a simple, rapid, and effective tool for detecting the A201S and F290V mutations in the ace-1 gene of S. frugiperda. The assay’s simplicity, isothermal operation, and clear visual results make it a valuable tool for resistance monitoring and integrated pest management in agricultural settings.
研究领域:
昆虫学
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分子生物学
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农药学
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农业害虫管理
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DOI
PubMed
高影响因子
FOOD CONTROL
·
2024
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IF 5.6
Nanfack V.D. Carole,Lina Sheng,Jian Ji,Shang Wu,Yinzhi Zhang,Xiulan Sun
Ensuring the safety and high quality of food relies on promptly and clearly identifying foodborne diseases, including Salmonella . Nucleic acid testing plays an important role in ensuring food safety. Commercially available high-throughput molecular extraction technologies in-field application is limited as they are reliant on specialized and expensive equipment. The 96-well microplate recycling through biopolymer inner surface coating, was confirmed with FTIR, XRD, and XPS. Once optimized the DNA extraction protocol was equipment-free, user and eco-friendly while reducing sample contamination. Using the biopolymer microplate, the extraction and direct LAMP amplification of Salmonella spp. from milk samples were assessed. The gDNA had an average 260/280 ratio of 1.97, with a DNA yield of 16.88 ± 5.40 μg·mL -1 from 9 × 10 10 CFU·mL -1 Salmonella . Subsequently, using acid chrome K we visually detected the presence of Salmonella . The quantity of bacteria was correlated to the increase in the blue intensity of the color indicator. A detection limit of 1.54 CFU·mL -1 was achieved with good specificity and sensitivity in an equipment free setting. This study successfully established a 96-well plate-based high-throughput equipment free DNA extraction method. The methods enable the high-throughput extraction and amplification of 96 samples concomitantly within 1hr 30 min corresponding respectively to a total of 1536 and 768 samples within a working day treated by a researcher without the use of specialized and expensive equipment. The polymeric modified microplate was used for high-throughput DNA extraction and direct molecular amplification for pathogen detection. High-throughput DNA extraction no longer requires cumbersome equipment.
研究领域:
食品安全
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分子生物学
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微生物学
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生物技术
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DOI
高影响因子
BIOSENSORS & BIOELECTRONICS
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2023
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IF 12.6
Hao Bai,Yuqing Liu,Linbo Gao,Tao Wang,Xiaoli Zhang,Jie Hu,Lisha Ding,Yueting Zhang,Qian Wang,Lei Wang,Jianlong Li,Zhifeng Zhang,Yang Wang,Chenlan Shen,Binwu Ying,Xiaoyu Niu,Wenchuang Hu
Screening for high-risk human papillomavirus (HPV) infection is one of the most important preventative measures for cervical cancer. However, fast, convenient, and low-cost HPV detection remains challenging, especially in resource-limited settings. Here, we report a portable all-in-one device (PAD) for point-of-care testing (POCT) for HPV16 and HPV18 DNA in cervical swabs. The PAD was engineered to integrate modules for extraction-free sample lysis, loop-mediated isothermal amplification (LAMP) with lyophilized reagent beads, and real-time colorimetric signal sensing into a single miniaturized device, considerably shortening the sample-to-result time to 15 min. The precision liquid handling in the completely sealed microfluidic chip is achieved by a uniquely designed pressure-balanced automatic liquid flow mechanism, thereby eliminating the need for manual manipulation of liquids and thus the risk of biohazards . The PAD employs an improved real-time colorimetric LAMP (rcLAMP) assay with a limit of detection (LOD) of 1 copy/μL, enabled by enhanced assay chemistry to maximize the reaction kinetics . To validate this device for clinical application, we tested 206 clinical cervical swab samples and obtained a sensitivity of 92.1% and a specificity of 99.0%. This custom PAD enabled by microfluidic and electronic engineering techniques can be configured for the simultaneous detection of HPV16 and HPV18 or other pathogens in point-of-care applications.
研究领域:
医疗设备
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分子生物学
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癌症研究
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即时诊断
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微流控技术
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2025
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IF 6.7
Yuchong Zheng,Wanjun Yao,Zerui Wu,Liqun He,Weidong Zheng,Zida Li
Accurate nucleic acid quantification is vital for clinical diagnostics, yet the widespread adoption of digital PCR remains limited due to its reliance on fluorescence detection and specialized microfluidics. We present Flow-LAMP, a label-free digital assay integrating loop-mediated isothermal amplification with scatter-based flow cytometric analysis of agarose gel beads. Polydisperse gel beads are formed by vortex emulsification and retain magnesium pyrophosphate precipitate in positive reactions. Flow cytometry enables volume and amplification readouts via forward (FSC) and side scattering (SSC) signals, respectively. We confirmed that SSC was strongly correlated with amplification products, while FSC-Height accurately reflected the bead volume. Using Epstein–Barr virus plasmid, Flow-LAMP achieved accurate quantification with a limit of detection of 38.15 copies/μL. Results from testing clinical plasma samples correlated well with qPCR and digital PCR. By eliminating fluorescent labeling and microfluidics, Flow-LAMP offers a cost-effective and accessible platform for digital nucleic acid detection using standard lab equipment.
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产品关联
DOI
PubMed
高影响因子
CLINICA CHIMICA ACTA
·
2025
·
IF 2.9
Yanghe Liu,Sijie Yin,Shiyue Chen,Chunlei Zhang,Nasifu Lubanga,Ping Tai,Mengqiu Xiong,Boyue Fan,Xincheng Yang,Xinyi Xia,PanFei Hou,Bangshun He
Helicobacter pylori ( H. pylori ) is a risk factor of gastrointestinal diseases. Vacuolating cytotoxin A (VacA) is a key component of the pathogenicity of H. pylori for gastric diseases , and detection for virulence genotypes facilitates precise clinical diagnosis and individual treatment. Hence, a more accurate, convenient, and highly sensitive detection method is urgently needed. In this study, we developed a method based on fluorescence-colorimetric dual-indicator loop-mediated isothermal amplification (FC-LAMP), allowing for naked-eye readout, using a constant temperature water bath within 40 min in clinical settings. Such an excellent sensitivity and specificity, and user-friendly operation approach achieved a limit of 10 −6 ng/μl for 16 s rRNA detection of H. pylori , and 100 % specificity for 55 clinical gastric fluid samples compared with the immunofluorescence staining (IFS) and quantitative polymerase chain reaction (qPCR) as reference methods. In short, this study offers a new strategy for detection of H. pylori and its virulence genotype in primary hospitals and in limited resources settings, facilitating individualized precision eradication of H. pylori .
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产品关联
DOI
PubMed
高影响因子
AUSTRALASIAN PLANT PATHOLOGY
·
2025
·
IF 1.1
Chen Jiajia,Huang Xiaoxiao,Peng Jinfeng,Chen Yuanyuan,Gafforov Yusufjon,Xing Yifan,Yu Jian
Globisporangium recalcitrans is an important phytopathogenic oomycete that causes soybean root rot. To develop a loop-mediated isothermal amplification (LAMP)-based system for rapid and specific detection of G. recalcitrans , four LAMP primers and two loop primers were designed with the Ribosomal DNA transcribed spacer 2 sequence (ITS2) as the target gene. The specificity and sensitivity of these primers were validated, and the system was also successfully applied to detect G. recalcitrans in soybean tissues after artificial inoculation and natural infection. The nucleic acid amplification reaction was performed under isothermal conditions at 63 °C for 60 min. Specificity was compared with those for 71 strains of G. recalcitrans , other Globisporangium spp. The minimum detection limit of the system was 100 fg·μL −1 for detecting fungal genomic DNA. This LAMP-based system provides a technique for specific detection of G. recalcitrans and rapid diagnosis of the disease it caused.
研究领域:
植物病理学
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分子生物学
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产品关联
DOI
高影响因子
MICROCHIMICA ACTA
·
2025
·Vol. 192
·期 12
·845
·
IF 5.3
Li Yirui,Sun Xiaodong,Gao Jiale,Wang Yan,Li Zhenghui,Niu Bing,Chen Qin
AuNFs (gold nanoflowers) are widely applied in the field of biosensing; their unique morphological characteristics and high specific surface area enable them to provide a large number of active sites, thereby enhancing the efficiency of catalytic reactions. These features make AuNFs a promising material for improving the performance of loop-mediated isothermal amplification (LAMP). Firstly, the reduction of 4-NP catalyzed by nano-gold (NG) with different morphologies showed that AuNFs made the yellow solution almost transparent in 4 min, similar to AuNPs (gold nanoparticles), with higher catalytic efficiency compared to AuNRs (gold nanorods) (> 8 min). The incorporation of AuNFs increased the activity of Bst DNA polymerase, and the kinetic parameters showed a V max / K m of 0.021, which was higher than that of AuNPs (0.015), AuNRs (0.013), and the LAMP system without NGs (0.013). Moreover, at ambient temperature, the UV–vis intensity of the characteristic peak indicates that the loading capacity of ssDNA is approximately 1.6 × 10 5 per AuNF, and when the temperature rises to 65 °C, ssDNA is almost completely released from the AuNFs. This mechanism may be beneficial for suppressing the generation of non-specific amplification. Dynamic light scattering (DLS) and circular dichronism (CD) results indicate that AuNFs alter the α-helix structure upon binding with Bst, potentially thereby enhancing enzyme activity. LAMP amplification was performed at different temperatures, and the results illustrate that the Tt values of AuNFs@LAMP were lower than those of AuNRs@LAMP ( P < 0.0001), AuNPs@LAMP ( P = 0.0082), and classical LAMP ( P < 0.0001) at 40, 45, 50, 55, and 60 ℃. Thus, we proposed the AuNFs@LAMP (AuNF-mediated LAMP) method and applied it to the detection of AHSV (African horse sickness virus) standard plasmid sample and spiked dust sample with a sensitivity as low as 10 copies/μL. Intra-day, inter-day, and intra-batch precision showed RSD < 5%, with RSD ranging from 2.51 to 8.75% across low, medium, and high plasmid template concentrations, respectively. Moreover, the specific AuNFs@LAMP system inhibits non-specific amplification in negative samples and in samples containing other equine virus plasmids (WNV, JEV, VSV, NIV, EEEV, WEEV). Graphical
研究领域:
纳米技术
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分子诊断
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病毒学
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生物传感器
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生物化学
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产品关联
DOI
PubMed
高影响因子
CROP PROTECTION
·
2021
·
IF 2.571
Ran Du,Tao Luo,Jing Zhang,Long Yang,Mingde Wu,Guoqing Li
The ascomycetous fungus Leptosphaeria biglobosa (Lb) and its sibling L. maculans (Lm) constitute a species complex responsible for blackleg of oilseed rape . L. biglobosa is composed of at least seven phylogenetic sub-clades, including L. biglobosa ‘brassicae’ (Lbb) and L. biglobosa ‘canadensis’ (Lbc), which widely distribute in America, Australia and Europe. So far, only Lbb has been reported in China, and recent surveys showed that Lbc widely occurred in Northern China. In order to track the spread of Lbc in this country, a simple and rapid diagnostic tool was developed based on loop-mediated isothermal amplification (LAMP) in this study. A total of 11 primer sets targeting the internal transcribed spacer region of the ribosomal DNA of Lbc were designed, and screened based on their performance in initiation of LAMP reactions for Lbc (65 °C, 40 min). The primer set PS5 with an introduced T-to-A mutation could efficiently and persistently detect Lbc. Then, specificity and sensitivity of PS5 was tested using the DNA from Lbb, Lbc, Lm, and 9 other fungi. The results showed that PS5 was specific to Lbc, and highly sensitive with the detection threshold of the Lbc DNA as low as 988 fg per reaction. The utility was evaluated using the DNA from healthy and Lbc-infected stems of oilseed rape, and the result showed that the LAMP assay positively detected the DNA from Lbc-infected stems. The LAMP technique developed herein will help to monitor populations of the blackleg pathogens and for developing strategies for management of the blackleg disease.
研究领域:
植物病理学
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分子生物学
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农业科学
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DOI
高影响因子
MICROCHEMICAL JOURNAL
·
2025
·
IF 5.1
Keyi Long,Tiao Han,Wenxi Hu,Fuping Nie,Mei Yang,Jun Li,Changjun Hou,Danqun Huo
Loop-mediated isothermal amplification (LAMP) has been used as one of the tools for nucleic acid detection. The pyrophosphoric acid (PPi) produced by LAMP leads to a change in the pH value of the solution, and acid-base indicators can be used to distinguish positive from negative. However, the microliter reaction system and low concentration target cause small changes in pH, sensitive pH indicators are urgently needed. Herein, we developed a one-step solvothermal approach to synthesize biomass-derived carbon dots named FA-bioCDs. Compared with hydrothermal synthesis, the solvothermal approach red-shifted the emission maximum to 535 nm, and made FA-CDs pH-sensitive in the range of 8.16–9.12, so it can be used for LAMP endpoint detection. The relatively alkaline LAMP buffer reduces the fluorescence of FA-bioCDs. Rapid LAMP accumulates hydrogen ions, lowering the pH and “turning on” FA-bioCDs fluorescence. Thus, LAMP acts as a pH-mediated fluorescence switch for FA-bioCDs. The strategy was used for human papillomavirus DNA detection, and the detection limit was as low as 2.450 copies·μL −1 in about 30 min. LAMP ensures rapid and sensitive detection, and the relatively long emission wavelength of FA-bioCDs may reduce interference in practical detection scenarios. The development of LAMP-FA-bioCDs not only offers a rapid and environmentally friendly tool for nucleic acid detection in clinical settings but also bridges the gap between agriculture and medicine.
研究领域:
纳米技术
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分子诊断
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生物分析化学
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基因工程
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即时检测
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产品关联
DOI
高影响因子
Coatings
·
2025
·Vol. 16
·期 1
·
IF 2.8
Yunxi Nie,Yushu Chen,Xinyou Liu
This study examines the pigment-dependent photoaging behavior of laboratory-prepared mock-up Chinese lacquer coatings colored with cinnabar, orpiment, and lapis lazuli under high-pressure mercury-lamp irradiation. Colorimetric results showed rapid changes within the first three days, with maximum ΔE values of 14.05 (red), 16.74 (yellow), and 17.97 (blue) after 30 days. Cinnabar-based films exhibited the highest color stability, whereas orpiment and lapis-lazuli coatings underwent pronounced hue shifts and chroma increases. Gloss loss and surface roughness evolution displayed a strong negative correlation: orpiment coatings experienced the most severe degradation, with gloss decreasing by over 60% and surface roughness increasing by approximately 70%, while cinnabar coatings showed the least decline (≈55% gloss loss; ≈27% roughness increase). SEM analysis further revealed extensive cracking and particle fragmentation in orpiment films, moderate surface disruption in lapis-lazuli films, and minimal microstructural damage in cinnabar films. Non-invasive reflection-mode FTIR spectroscopy confirmed these trends, showing minimal chemical change in cinnabar coatings but significant carbonyl growth, C–O–C band broadening, and aliphatic chain cleavage in orpiment and lapis-lazuli coatings. These results highlight the critical role of pigment chemistry in modulating UV-induced degradation pathways. Integrating optical, morphological, and chemical evidence, this study establishes a clear pigment-dependent degradation mechanism and provides valuable guidance for evaluating the long-term stability of lacquered cultural heritage and optimizing modern lacquer formulations.
研究领域:
文化遗产科学
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聚合物降解
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材料化学
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文物保护科学
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表面化学
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DOI
高影响因子
PLANT DISEASE
·
2018
·Vol. 102
·期 8
·
IF 2.941
F. Fan,W. X. Yin,G. Q. Li,Y. Lin,C. X. Luo
Resistance to succinate dehydrogenase inhibitors (SDHI) in Botrytis cinerea is associated with point mutations in the target gene succinate dehydrogenase subunit B ( SdhB ). The substitution from histidine to arginine at codon 272 (H272R) is currently the predominant mutation in SDHI-resistant populations in B. cinerea worldwide. In order to monitor the development of resistance to SDHI, a rapid, simple, and efficient method with high specificity to the H272R point mutation was developed based on loop-mediated isothermal amplification (LAMP). To specifically detect the H272R mutation, a set of four primers was designed based on the sequence of SdhB , and the LAMP reaction was optimized. When SYBR Green I was added after reaction, only samples with the H272R mutation showed the color change (from brown to fluorescent yellow), indicating that this set of primers could successfully discriminate the H272R genotype from other genotypes. Specificity and accuracy tests showed that this LAMP assay had high specificity and accuracy. Moreover, the LAMP method was further simplified with fungal mycelia and conidia as the amplification template which could be prepared within 5 min. Due to the low cost, simplicity, high efficiency, and specificity, the developed LAMP assay may contribute to the monitoring of resistance development to SDHI in B. cinerea , especially in field and high-throughput experiments.
研究领域:
植物病理学
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分子生物学
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农药抗性
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真菌遗传学
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检测技术
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DOI
高影响因子
危险品化学品经营许可证（带存储）