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rapid detection of procalcitonin using dual-color carbon quantum dots as fluorescent probes in FRET assay

SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY 2025年 338卷 126207页

作者： Tian, Yuxin Du, Yuwei Chen, Zhang Li, Li Yan, Ruhong Li, Xiaoqiang Yue, Juan Chinese Acad Sci Suzhou Inst Biomed Engn & Technol CAS Key Lab Biomed Diagnost Suzhou 215163 Peoples R China Nanjing Univ Suzhou Hosp Affiliated Hosp Suzhou Res CtrMed Sch Suzhou 215153 Peoples R China Univ Sci & Technol China Sch Biomed Engn Suzhou Div Life Sci & Med Hefei 230026 Peoples R China

FRET, known as Forster Resonance Energy Transfer, serves as a versatile technique enabling precise quantitative assessment of protein concentrations with minimal disruption to biological processes. In this study, we have developed two types of fluorescent carbon quantum dots, B-CDs and G-CDs, serving as donor and acceptor probes, respectively, for FRET-based procalcitonin (PCT) detection. The B-CDs exhibit strong blue fluorescence (excitation/emission peaks at 360/442 nm) as donors, while the G-CDs display faint green fluorescence (excitation/emission peaks at 449/520 nm) as acceptors. These CDs are conjugated with PCT antibodies to form immune probes. Upon antigen binding, the antibody quickly binds and reacts with the antigen. Concurrently, the fluorescence energy from the donor is transferred to the acceptor, leading to a decrease in the blue fluorescence intensity emitted by B-CDs. The ratio of the fluorescence intensity between the green probe (G-CDs-Ab2) and the blue probe (B-CDs-Ab1) exhibited a positive correlation with the concentration of PCT antigen in the low concentration range of 0.5 to 5 ng/mL. Consistent results were observed in both phosphate-buffered saline (PBS) and diluted human serum. In the concentration range of 0.5-5 ng/mL in PBS buffer, the linear equation is expressed as FA/FD _-0.0045 x c [PCT]<^>2 + 0.0021 x c [PCT] + 0.3232, with a limit of detection (LOD) of 0.1 ng/mL based on 3 sigma K-1. For concentrations spanning from 5-80 ng/mL, the equation is FA/FD = 0.0021 x c [PCT] + 0.4212. These findings suggest that carbon dot-labeled immune probes could offer a novel approach for sensitive detection of disease biomarkers.

关键词： FRET Carbon dots Fluorescent Procalcitonin rapid detection Dual-color

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rapid detection of Staphylococcus aureus in animal-derived foods using aptamer-based fluorescent lateral flow biosensors

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BIOCHEMICAL ENGINEERING JOURNAL 2025年 220卷

作者： Zhou, Shuxian Wang, Yinghui Wang, Haifang Dou, Shouyi Liu, Mengyue Dong, Haowei Li, Zhengtao Li, Donghan Liu, Jingjing Sun, Xia Guo, Yemin Darwish, Ibrahim A. Wu, Yanfang Shandong Univ Technol Coll Agr Engn & Food Sci 266 Xincun Xilu Zibo 255049 Shandong Peoples R China Shandong Prov Engn Res Ctr Vegetable Safety & Qual 266 Xincun Xilu Zibo 255049 Shandong Peoples R China Beijing Univ Chinese Med Dongzhimen Hosp Beijing 100700 Peoples R China Dezhou Univ Inst Biophys Shandong Prov Key Lab Biophys Dezhou 253023 Shandong Peoples R China King Saud Univ Coll Pharm Dept Pharmaceut Chem POB 2457 Riyadh 11451 Saudi Arabia Univ Otago Dept Chem Dunedin 9016 New Zealand

Staphylococcus aureus (S. aureus), commonly found in animal-derived foods, poses a significant threat to food safety. Traditional detection methods often suffer from limited sensitivity and selectivity, are time-consuming, and involve complex procedures. In this study, fluorescent lateral flow biosensors (LFB) based on aptamer recognition were developed for the rapid and cost-effective detection of S. aureus. The method utilizes a Cy5labeled aptamer (Cy5-Apt) as a signal recognition molecule. detection is achieved through the competitive binding of S. aureus and a complementary strand to the aptamer, enabling the fluorescence intensity measurement on the LFB within 10 min. Key factors including the Cy5-Apt concentration, the length of the linker arm, the position and length of the complementary strand to the aptamer, BSA concentration, and buffer composition were optimized. The method showed good linear response for the quantitative detection of S. aureus in the concentration range of 2.8 x 101 to 2.8 x 107 CFU mL-1, and the limit of detection (LOD) was 1.65 CFU mL-1. Application tests in milk and chicken meat confirmed the selectivity and sensitivity of this enzyme-and antibody-free approach. This aptamer-based method offers a promising solution for low-cost, rapid, and efficient detection of S. aureus in food safety applications.

关键词： Aptamer Fluorescent lateral flow biosensors Staphylococcus aureus rapid detection Food safety

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rapid detection of sludge moisture content based on the frequency domain reflection

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ENVIRONMENTAL TECHNOLOGY 2025年 1-14页

作者： Zhang, Yan Yao, Yawen Yu, Shuying Huang, Minrui Lu, Xilong Lian, Jiadi Xu, Peng Rao, Binqi China Jiliang Univ Coll Mech & Elect Engn Hangzhou 310018 Peoples R China Zhejiang Crown Power Tools Manu Co Ltd Jinhua Peoples R China Zhejiang Hongshi Environm Protect Co Ltd Jinhua Peoples R China China Jiliang Univ Coll Energy Environm & Safety Engn Hangzhou Peoples R China

The moisture content (MC) of sludge is a critical factor affecting sludge treatment and disposal technologies. Traditional drying methods for measuring sludge MC are time-consuming and lack of portability. To achieve rapid online detection of sludge MC, this paper proposes a method based on frequency domain reflectometry. The theoretical feasibility of detecting MC using the frequency reflection principle was demonstrated. The model of the relationship between MC and dielectric constant, as well as the relationship between dielectric constant and oscillation frequency, was derived. The mathematical model correlating MC of sludge with oscillation frequency was established, and the structure of the sensor probe was designed. Experimental results show that the third-degree fitting curve achieving the best $R_t<^>2$Rt2 of 0.98247. The proposed method presents good measurement performance compared to the drying method, with satisfactory repeatability and consistency in performance tests. For the sludge samples with MC ranging from 10% to 75%, the maximum error is 3.4%, and the average absolute error is 1.86%. This method provides an effective way for rapid detection of sludge MC, offering shorter measurement times and better portability compared to traditional methods.

关键词： Frequency domain reflectometry sludge moisture content detection oscillation frequency rapid detection

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rapid detection of Heavy Metal Ions Co³⁺, Cu²⁺, Ni²⁺, and Zn²⁺ in Wastewater By Pyrimidine-Thiourea Ligand

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JOURNAL OF APPLIED SPECTROSCOPY 2024年第2期91卷 428-433页

作者： Wang, Zhou Jin, Feng Panzhihua Univ Coll Vanadium & Titanium Panzhihua Peoples R China Anhui Univ Sci & Technol Med Coll Huainan Peoples R China

rapid detection of trace heavy metal ions in water is of great relevance to the environment and human health in the future. With pyrimidine as the parent structure, the water-soluble thiourea group is skillfully introduced to form a symmetric framework, thus firmly capturing the heavy metal ions in the water. The responsiveness of heavy metal ions in water was observed by UV-fluorescence spectroscopy. Especially with Co(NO3)(2), Cu(NO3)(2), Ni(NO3)(2), and Zn(NO3)(2), the response changes are most significant. The rapid response changes between them and trace heavy metal ions in water were intuitively observed under sunlight and fluorescence irradiation environment.

关键词： rapid detection high sensitivity photophysical characteristics pyrimidine-thiourea ligand

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rapid detection of water content in donkey-hide gelatin based on terahertz spectroscopy and effective medium theory

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JOURNAL OF FOOD MEASUREMENT AND CHARACTERIZATION 2024年第10期18卷 8135-8144页

作者： Liu, Lingyu Wu, Hao Wang, Zhongmin Li, Yizhang Li, Ke Guo, Yongbin Yang, Xiuwei Qilu Univ Technol Inst Automat Shandong Acad Sci 19 Keyuan Rd Jinan 250014 Peoples R China Beijing Microelect Technol Inst Beijing Peoples R China Capital Normal Univ Dept Phys Beijing 100048 Peoples R China

In this study, a rapid water content detection method for donkey-hide gelatin was established based on terahertz time-domain spectroscopy in reflection mode. Notably, the effective medium theory, informed by the double-Debye relaxation for free water, was employed to predict the water content in the sample. The water content obtained through effective medium theory was lower than that of the traditional thermogravimetric method due to the existence of bound water. Bound water is invisible in THz spectroscopy due to its slower dynamics. In the frequency range of 0.22-0.48 THz, the offset in measured values between the thermogravimetric method and the THz-based method was fitted by a polynomial function. The results of THz-based method calibrated by polynomial fitting were consistent with the findings of the thermogravimetric method. The water content detection results derived by these two methods demonstrated high correlation values (R2) of 0.9860, 0.9160, and 0.9784, and small root mean square errors (RMSE) of 4.5584, 4.4887, and 5.3657 for three samples, respectively. Moreover, the proposed water content detection method needs to be calibrated only once by the thermogravimetric method. Overall, this study provides a rapid water content detection method and demonstrates the potential of real-time water content detection in donkey-hide gelatin.

关键词： rapid detection Water content Terahertz Effective medium theory Donkey-hide gelatin

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rapid detection of Salmonella typhimurium in food samples using electrochemical sensor

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LWT-FOOD SCIENCE AND TECHNOLOGY 2024年 206卷

作者： Wang, Yimin He, Xun Wang, Shan Ma, Jinyou Hu, Dongfang Liang, Haiyang Ma, Caoyuan Jin, Yafeng Chen, Xuanqi Xu, Guangri Wang, Lei Henan Inst Sci & Technol Coll Anim Sci & Vet Med Xinxiang 453003 Henan Peoples R China Minist Educ Key Lab Anim Pathogens & Biosafety Zhengzhou Peoples R China Henan Inst Sci & Technol Sch Chem & Chem Engn Xinxiang 453003 Henan Peoples R China

rapid and accurate detection of pathogenic bacteria is crucial for ensuring food safety and public health. In this study, we aimed to develop a novel molecularly imprinted polymer sensor based on screen-printed electrodes for the specific detection of Salmonella typhimurium. The sensor was constructed by electropolymerizing dopamine in the presence of Salmonella typhimurium on the electrode surface, followed by the removal of the bacteria to create specific binding cavities. The sensor demonstrated excellent specificity for Salmonella typhimurium, with a detection limit approximately of 101 CFU/ml and a detection time of only 4 min, with the sensor accurately differentiating Salmonella typhimurium from other common foodborne pathogens such as Escherichia coli and Listeria monocytogenes. The performance of the sensor was validated using real food samples, including pork and milk, showing its well suited for rapid and on-site detection of pathogenic bacteria. The development of this molecularly imprinted polymer with electrochemical sensor represents a significant advancement in the field of electrochemical biosensors, offering a promising tool for food safety monitoring and public health protection.

关键词： Salmonella typhimurium Electrochemical sensor Molecularly imprinted polymer Food safety rapid detection

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rapid detection of Aeromonas veronii Using GGDEF Gene-Based Colorimetric Loop-Mediated Isothermal Amplification

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JOURNAL OF FISH DISEASES 2025年第0期 e14112页

作者： Lau, Tien Tien Vicky Puah, Suat Moi Chua, Kek Heng Univ Malaya Fac Med Dept Biomed Sci Kuala Lumpur Malaysia

Aeromonas veronii has emerged as an important fish pathogen that poses threats to the aquaculture industry worldwide, and rapid detection is essential to minimise negative economic impact. In this study, a colorimetric loop-mediated isothermal amplification (LAMP) assay was developed to rapidly detect A. veronii, targeting a gene encoding GGDEF domain-containing diguanylate cyclase. The entire LAMP reaction could be completed in 30 min at 65 degrees C based on colour change from pink to yellow. In addition, the assay demonstrated 100% specificity with no cross reaction with other common fish pathogens. The detection limit (LOD) was 500 pg/mu L using purified plasmid and 1.72 x 10(5) cfu/mL (equivalent to 1.72 x 10(2) cfu/reaction) using crude genomic DNA extracted from the pure culture of A. veronii ATCC 9071. LAMP demonstrated comparable performance to conventional PCR using 57 bacterial isolates, with 100% (20/20) specificity, 91.9% (34/37) sensitivity, 94.7% (54/57) accuracy and a 0.888 Cohen's kappa value. Lastly, the LOD of LAMP in a spiked water sample was 1.72 x 10(5) cfu/mL (equivalent to 3.44 x 10(3) cfu/reaction). Overall, our LAMP assay has a high level of diagnostic agreement with conventional PCR and can be used as a valuable tool for the rapid detection of A. veronii from environmental samples.

关键词： Aeromonas veronii colorimetric loop-mediated isothermal amplification (LAMP) fish pathogen rapid detection sensitivity specificity

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rapid detection of Pan-Avian Influenza Virus and H5, H7, H9 subtypes of Avian Influenza Virus using CRISPR/Cas13a and lateral flow assay

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POULTRY SCIENCE 2025年第2期104卷 104745页

作者： Yang, Yujia Yang, Zhiyi Zhang, Xinkui Niu, Beibei Huang, Qiuhong Li, Yan Yin, Huifang Zhang, Xianpeng Liao, Ming Jia, Weixin South China Agr Univ Coll Vet Med Guangdong Engn Lab Medicament Zoonosis Prevent & C Key Lab Zoonoses Prevent & Control Guangdong Prov Guangzhou 510642 Peoples R China Longyan Univ Fujian Prov Univ Coll Life Sci Engn Res Ctr Prevent & Control Anim Original Zoono Longyan 364012 Peoples R China Dongguan Ctr Anim Dis Prevent & Control Dongguan Key Lab Zoonosis Dongguan 523128 Peoples R China

Avian Influenza Virus (AIV) has been prevalent worldwide in recent years, resulting in substantial economic losses in the poultry industry. More importantly, AIV is capable of cross-species transmission among mammals, posing a dormant yet considerable threat to human health and safety. In this study, two rapid detection methods for AIV based on the CRISPR-Cas13a were developed. These methods can identify AIV through the M gene and differentiate the H5, H7, and H9 subtypes via the HA gene. The first method utilizes RT-RAA isothermal amplification of the target sequence in combination with the "collateral effect" of the Cas13a protein. The results are measured using a real-time quantitative PCR instrument, with a Limit of detection (LOD) as low as 1 copy/ mu L. The second method combines RT-RAA with Cas13a and a lateral flow assay, allowing results to be visually observed with the naked eye, with a LOD of 10 copies/mu L. Both methods demonstrated specificity and sensitivity comparable to or exceeding that of qRT-PCR, suggesting strong potential for clinical application.

关键词： Avian influenza virus rapid detection CRISPR/Cas13a RT-RAA Lateral flow assay

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rapid detection of non-normal teeth on dental X-ray images using improved Mask R-CNN with attention mechanism

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INTERNATIONAL JOURNAL OF COMPUTER ASSISTED RADIOLOGY AND SURGERY 2024年第4期19卷 779-790页

作者： Guo, Yanbin Guo, Jing Li, Yong Zhang, Peng Zhao, Yuan-Di Qiao, Yundi Liu, Benyuan Wang, Guoping Huazhong Univ Sci & Technol Coll Life Sci & Technol Dept Biomed Engn Hubei Bioinformat & Mol Imaging Key Lab Wuhan 430074 Peoples R China Nanchang Univ Affiliated Stomatol HospJiangxi Prov Clin Res Ctr Jiangxi Med CollJiangxi Prov Key Lab Oral Biomed Jiangxi Prov Clin Res Ctr Oral DisDept Dent Gen & Nanchang 330038 Jiangxi Provinc Peoples R China Nanchang Univ Affiliated Hosp 1 Dept Anesthesiol Nanchang 330006 Peoples R China Univ Massachusetts Lowell Dept Comp Sci Lowell MA 01854 USA

PurposeDental health has been getting increased attention. Timely detection of non-normal teeth (caries, residual root, retainer, teeth filling, etc.) is of great importance for people's health, well-being, and quality of life. This work proposes a rapid detection of non-normal teeth based on improved Mask R-CNN, aiming to achieve comprehensive screening of non-normal teeth on dental X-ray *** improved Mask R-CNN based on attention mechanism was used to develop a non-normal teeth detection method trained on a high-quality annotated dataset, which can segment the whole mask of each non-normal tooth on the dental X-ray image *** average precision (AP) of the proposed non-normal teeth detection was 0.795 with an intersection-over-union of 0.5 and max detections (maxDets) of 32, which was higher than that of the typical Mask R-CNN method (AP = 0.750). In addition, validation experiments showed that the evaluation metrics (AP, recall, precision-recall (P-R) curve) of the proposed method were superior to those of the Mask R-CNN method. Furthermore, the experimental results indicated that proposed method exhibited a high sensitivity (95.65%) in detecting secondary caries. The proposed method took about 0.12 s to segment non-normal teeth on one dental X-ray image using the laptop (8G memory, NVIDIA RTX 3060 graphics processing unit), which was much faster than conventional manual *** proposed method enhances the accuracy and efficiency of abnormal tooth diagnosis for practitioners, while also facilitating early detection and treatment of dental caries to substantially lower patient costs. Additionally, it can enable rapid and objective evaluation of student performance in dental examinations.

关键词： rapid detection Non-normal teeth Improved Mask R-CNN Attention mechanism X-ray images

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rapid detection of Staphylococcus aureus in blood culture samples using human IgG-based lateral flow assay

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MICROBIOLOGY SPECTRUM 2024年第2期12卷

作者： Srisrattakarn, Arpasiri Charoensri, Nicha Prompipak, Jeerati Ouancharee, Wajeeorn Saiboonjan, Bhanubong Tippayawat, Patcharaporn Chanawong, Aroonwadee Wonglakorn, Lumyai Kanwattanee, Ekgarak Piyapatthanakul, Sirikan Masmalai, Thitimar Ariyapim, Anisara Kendal, Rinjong Promson Lulitanond, Aroonlug Khon Kaen Univ Fac Associated Med Sci Ctr Res & Dev Med Diagnost Labs Khon Kaen Thailand Khon Kaen Univ Fac Associated Med Sci Ctr Innovat & Stand Med Technol & Phys Therapy Khon Kaen Thailand Khon Kaen Univ Srinagarind Hosp Clin Microbiol Unit Khon Kaen Thailand Queen Sirikit Natl Inst Child Hlth Clin Microbiol Lab Bangkok Thailand Univ Queen Sirikit Heart Ctr Northeast Clin Lab Khon Kaen Thailand Khon Kaen Hosp Khon Kaen Thailand

Staphylococcus aureus is one of the most common pathogens. The conventional workflow for identifying this organism is time-consuming and takes up to several days. Therefore, we developed a colloidal gold-based lateral flow immunoassay (LFIA) using human IgG as a conjugated antibody to detect S. aureus. One hundred and thirty-eight clinical isolates, including 79 S. aureus and 59 non-S. aureus were spiked in blood samples, and incubated at 37degree celsius for 24 h. The bacterial antigens were simply extracted before being tested by the developed LFIA strips. The results were read by the naked eye within 15 min. Conventional PCR was used as a reference method. The sensitivity and specificity of the developed LFIA were 100% (95% CI: 94.2%-100.0% and 92.4%-100.0%, respectively) in spiked blood culture samples. The detection limits of the LFIA for the purified protein A and bacterial colonies were 10(-3) mu g/mL and 10(7) CFU/mL, respectively. The performance of the LFIA testing in 221 bacterial colony isolates and 118 positive blood culture bottles from three hospitals by their medical technologists showed 98.1% (95% CI: 94.1%-99.5%) and 89.7% (95% CI: 79.3%-95.4%) sensitivity, respectively. The LFIA is a quick, easy, and sensitive method for detecting S. aureus without expensive equipment. It might have the potential for early diagnosis of routine service in low-resource laboratories, leading to a rapid and effective treatment. IMPORTANCE In this study, we modified our previously developed lateral flow immunoassay (LFIA) test for the detection of Staphylococcus aureus by using an in-house human IgG as a conjugated antibody instead of the specific commercial antibody. It gave comparable results to the former developed-LFIA test and helped cost reduction.

关键词： lateral flow immunoassay positive blood culture samples rapid detection Staphylococcus aureus

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