ANALYTICAL CHEMISTRY
·
2025
·
IF 6.7
Junzhuo Shan,Yixiao Sheng,Lun Luo,Wenhai Wang,Xinjiang Liu,Yi Ma,Jufang Wang
Apurinic/apyrimidinic endonuclease 1 (APE1) is a key enzyme involved in DNA repair and cellular redox regulation, and is frequently overexpressed in tumor cells. This highlights the urgent need for a rapid and high-sensitive point-of-care testing (POCT) strategy for APE1 to facilitate early cancer diagnosis. Rolling circle amplification (RCA) is a widely used isothermal DNA amplification method; however, its application in APE1 detection remains rare. Here, we introduce a versatile RCA-Lock (Rlock) conversion platform that enables the transformation of RCA-based nucleic acid detection technologies into APE1-responsive assays. Building upon this platform, we further developed a novel POCT method for APE1 detection─Rlock-mediated, CRISPR/Cas12a-driven RCA cycle (RCRE)─which, for the first time, integrates CRISPR/Cas12a with RCA into a one-pot APE1 detection system. The RCRE assay achieves a limit of detection of 8.86 × 10–4 U/mL within 30 min, while requiring minimal equipment, low cost, and no complex handling procedures. This Rlock-based conversion strategy represents a transformative advance in the field of APE1 diagnostics and offers conceptual inspiration for the design of programmable nucleic acid–based biosensors. The resulting RCRE assay significantly broadens the technological landscape for early cancer detection and paves the way for future clinical translation.
研究领域:
分子诊断学
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生物化学
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生物技术
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癌症研究
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遗传学
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分析化学
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产品关联
DOI
PubMed
高影响因子
ANALYST
·
2021
·Vol. 146
·期 22
·6960-6969
·
IF 4.616
Ping Mao,Lei Cao,Zedong Li,Minli You,Bin Gao,Xianghong Xie,Zhenrui Xue,Ping Peng,Chunyan Yao,Feng Xu
Digital PCR has shown great potential for quantitative nucleic acid testing (NAT), but most existing platforms are dependent on large auxiliary equipment (e.g., vacuum pump, amplification instrument, fluorescence microscope) to achieve target dispersion, amplification, signal capture and result analysis. Such complex, expensive and bulky NAT platforms have limited their applications in resource-limited areas, especially for point-of-care testing (POCT). In this work, we designed a digital isothermal NAT platform based on a pump-free open droplet array microfluidic chip. A pump-free microfluidic chip was developed based on an open microdroplet array in the form of thousands of independent microdroplets for spontaneous sample dispersion, without the need for external power. Combined with a handheld fluorescent signal reader based on a smartphone, this digital NAT platform can accurately quantify as low as 1 copy per μL of λDNA. Therefore, our integrated NAT platform, as a potable, robust and low-cost tool for highly accurate NA quantitative analysis, holds great potential for POCT applications.
研究领域:
分子生物学
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生物医学工程
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医学诊断
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DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2021
·
IF 10.618
Jiang Liu,Wei Lu,Lu Zhang,Jiao Yang,Zhong-Ping Yao,Yongcheng He,Yingchun Li
Electrochemical sensors have shown great advantage and application potential in point-of-care testing (POCT) related scenarios. However, some fatal problems plague its widespread utilization, which include the susceptibility of sensors to interference in real samples (e.g. pH), the contradiction between the limited objects detectable for most sensors and the requirement of multi-target analysis in most cases, and the complicated procedures in sensor preparation as well as in routine use. This paper contributed a tip-like electrochemical sensor prototype. By integrated with a commercial pipettor, it fulfilled semi-automatic assay procedure of sampling, detection and rinsing, thus saving operational time and manual work. The tip sensor owns the property of simple fabrication and is free from any modification of extra bio/chem materials. Moreover, built on multiple electrochemical signal outputs including open circuit potential , peak current and potential of specific electrochemical reaction , this work established a novel multi-component sensing strategy, wherein detection of uric acid (UA), urea and pH in urine samples was realized by using one single working electrode . The detection range for the above targets is 5.0~600 μM for UA, 4.0~8.0 for pH and 0.5~7.0 mM for urea with the detection limits ( S / N = 3) of 0.05 μM for UA and 5.4 μM for urea, and the sensitivity of pH assay is 73 mV/pH. Notably, as variation of sample pH has impact on electrochemical analysis , the pH-related parameter was introduced for calibration to diminish such interference. The developed tip sensor and the novel sensing strategy may open a new window for electrochemical technology and broaden its application in POCT.
研究领域:
电化学
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医学诊断
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生物传感器
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产品关联
DOI
PubMed
高影响因子
COLLOIDS AND SURFACES B-BIOINTERFACES
·
2022
·
IF 5.999
Laibao Zheng,Wenjia Dong,Chaochuan Zheng,Yunqiu Shen,Ruolan Zhou,Zhenxing Wei,Zhixuan Chen,Yongliang Lou
Rapid and portable detection of foodborne pathogens is of great significance for food safety and public health. The colorimetric methods based on naked-eye have been demonstrated to be a suitable qualitative method for point-of-care testing (POCT). However, analytical instruments like a microplate reader must be needed for the quantitative assay. To overcome its limitation, we herein report a novel photothermal method for foodborne pathogens based on the photothermal effect of aggregated mercaptophenylboronic acid-functionalized AuNPs (MPBA-AuNPs) induced by MPBA to translate the colorimetric detection into a simple temperature measurement using thermometers as the readout. The aggregated AuNPs show higher photothermal conversion efficiency than well-separated AuNPs under 660 nm laser irradiation. In the presence of bacteria, MPBA-AuNPs will attach to the surface of bacteria and keep separated from aggregation induced by excess MPBA, resulting in a lower temperature increase under 660 nm laser irradiation. Using E. coli O157:H7 as a model target, a good linear relationship is observed between temperature increase and bacteria concentration from 1.00 × 10 5 -1.00 × 10 9 cfu mL −1 (R 2 = 0.9877) with a detection limit of 1.97 × 10 4 cfu mL −1 , which is three orders of magnitude lower than of the MPBA-AuNPs-based colorimetric assays . The proposed photothermal method provided a universal platform for rapid and portable detection of broad-spectrum bacteria strains in real samples.
研究领域:
食品安全
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生物医学工程
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分析化学
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公共卫生
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DOI
高影响因子
Journal of Pharmaceutical Analysis
·
2024
·
IF 6.1
Yue Liu,Taimei Cai,Sen Chen,Tao Wen,Hailong Peng
The overuse of antibiotics has led to the severe contamination of water bodies, posing a considerable hazard to human health. Therefore, the development of an accurate and rapid point-of-care testing (POCT) platform for the quantitative detection of antibiotics is necessary. In this study, Cerium oxide (CeO 2 ) and Ferrosoferric oxide (Fe 3 O 4 ) nanoparticles were simultaneously encapsulated into N-doped nanofibrous carbon microspheres to form of a novel nanozyme (CeFe-NCMzyme) with a porous structure, high surface area, and N-doped carbon material properties, leading to a considerable enhancement of the peroxidase (POD)-like activity compared with that of the CeO 2 or Fe 3 O 4 nanoparticles alone. The POD-like activity of CeFe-NCMzyme can be quenched using L-Cysteine (Cys) and subsequently restored by the addition of a quinolone antibiotic (norfloxacin, NOR). Therefore, CeFe-NCMzyme was used as a colorimetric sensor to detect NOR via an “On-Off” model of POD-like activity. The sensor possessed a wide linear range of 0.05-20.0 μM ( R 2 = 0.9910) with a detection limit of 35.70 nM. Furthermore, a smartphone-assisted POCT platform with CeFe-NCMzyme was fabricated for quantitative detection of NOR based on RGB analysis. With the use of the POCT platform, a linear range of 0.1-20.0 μM and a detection limit of 54.10 nM were obtained. The spiked recoveries in the water samples were ranged from 97.73% to 102.01%, and the sensor exhibited good accuracy and acceptable reliability. This study provides a portable POCT platform for the on-site and quantitative monitoring of quinolone antibiotics in real samples, particularly in resource-constrained settings.
研究领域:
环境科学
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分析化学
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生物技术
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即时诊断
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2023
·
IF 7.4
Liping Xia,Juxin Yin,Jianjian Zhuang,Weihong Yin,Zheyu Zou,Ying Mu
Rapid and sensitive pathogen detection methods are critical for disease diagnosis and treatment. RPA-CRISPR/Cas12 systems have displayed remarkable potential in pathogen detection. A self-priming digital PCR chip is a powerful and attractive tool for nucleic detection. However, the application of the RPA-CRISPR/Cas12 system to the self-priming chip still has great challenges due to the problems of protein adsorption and two-step detection mode of RPA-CRISPR/Cas12. In this study, an adsorption-free self-priming digital chip was developed and a direct digital dual-crRNAs (3D) assay was established based on the chip for ultrasensitive detection of pathogens. This 3D assay combined the advantages of rapid amplification of RPA, specific cleavage of Cas12a, accurate quantification of digital PCR, and point-of-care testing (POCT) of microfluidics, enabling accurate and reliable digital absolute quantification of Salmonella in POCT. Our method can provide a good linear relationship of Salmonella detection in the range from 2.58 × 101 to 2.58 × 104 cells/mL with a limit of detection ∼0.2 cells/mL within 30 min in a digital chip by targeting the invA gene of Salmonella. Moreover, the assay could directly detect Salmonella in milk without nucleic acid extraction. Therefore, the 3D assay has the significant potential to provide accurate and rapid pathogen detection in POCT. This study provides a powerful nucleic detection platform and facilitates the application of CRISPR/Cas-assisted detection and microfluidic chips.
研究领域:
分子生物学
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生物技术
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微生物学
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分析化学
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即时诊断
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产品关联
DOI
PubMed
高影响因子
Small Methods
·
2024
·
IF 10.7
Chunyang Li,Yujian Li,Shaoyan Wu,Gui Li,Juan Li,Yan Zhao,Huan Cai,Jia Shu,Mingxuan Song,Qing Fu,Jianbo Yuan,Xin Gao,Zhujun Ai,Xiaosong Li,Rui Chen,Zhong Zuo
Catalytic nanoparticle@metal-organic framework (MOF) composites have attracted significant interest in point-of-care testing (POCT) owing to their prominent catalytic activity. However, the trade-off between high loading efficiency and high catalytic activity remains challenging because high concentrations of nanoparticles tend to cause the misjoining and collapse of the MOFs. Herein, a facile strategy is reported to encapsulate high concentrations of platinum (Pt) nanoparticles into zeolitic imidazolate framework-8 (ZIF-8) using polydopamine (PDA) as a support for Pt@ZIF-8 and as a flexible scaffold for further immobilization of Pt nanoparticles. The resulting composite (Pt@ZIF-8@PDA@Pt) exhibits ultrahigh Pt nanoparticle loading efficiency, exceptional catalytic activity, stability, and a bright colorimetric signal. Following integration with lateral flow immunoassay (LFIA), the detection limits for pre- and post-catalysis detection of B-type natriuretic peptide (NT-proBNP) are 0.18 and 0.015 ng mL −1 , respectively, representing a 6-fold and 70-fold improvement compared to gold nanoparticle-based LFIA. Moreover, Pt@ZIF-8@PDA@Pt-based LFIA achieves 100% diagnostic sensitivity for NT-proBNP in a cohort of 184 clinical samples.
研究领域:
纳米技术
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催化
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诊断
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材料科学
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生物医学工程
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产品关联
DOI
PubMed
高影响因子
Aggregate
·
2024
·
IF 13.9
Yi Long,Shurui Tao,Dongni Shi,Xingyu Jiang,Tian Yu,Yingxi Long,Libing Song,Guozhen Liu
Developing the sensitive point-of-care testing (POCT) of oncogenic nucleic acids from human papillomavirus (HPV) infection is essential in preventing cervical cancer, especially in resource-limited settings. Rolling circle amplification (RCA) is attractive in achieving POCT via nucleic acid-based aggregation under isothermal conditions. However, the influence of RCA product structure on the aggregation remains unexplored resulting in limited sensitivity. Here, a minimum secondary structured RCA technique (MSS-RCA) is developed by designing a unique circular template, demonstrating significantly enhanced detection sensitivity with only one amplification step and one primer under isothermal conditions. The amplification efficiency of MSS-RCA could be kinetically manipulated by controlling the secondary structure of the circular template. Introducing the invertase probe to MSS-RCA, HPV16 E6/E7 nucleic acid target was detected with a personal glucose meter (PGM) with a sensitivity of 5 f m (50 zmol in 10 µL). This integrated MSS-RCA-PGM detection system was successfully applied to detect HPV16 E6/E7 mRNA extracted from 54 cervical swab samples reaching a positive predictive value of 100.00% and negative predictive values of 96.00% (77.77% to 99.40%, 95% CI). MSS-RCA-PGM provides a sensitive POCT platform for the detection of nucleic acid biomarkers for screening of cervical cancer or other diseases.
研究领域:
分子生物学
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癌症筛查
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即时诊断
·
核酸扩增
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产品关联
DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2023
·
IF 8.4
Jiaxin Peng,Xiaomin Hang,Huiyi Wang,Kairen Zhao,Honghong Wang,Haoming Yu,Li Wang
MicroRNAs (miRNAs) have emerged as powerful biomarkers for various diseases diagnosis, yet their clinical application is greatly plagued by the lack of reliable and rapid detection methods. Here, a simple, fast, and accurate point-of-care testing (POCT) platform for miRNA-155 detection was developed by interfacing self-primer exponential amplification reaction (SPEXPAR) with colorimetric and POCT lateral flow assay (LFA). In this platform, a hairpin template was designed for exponential amplification reaction, which could effectively avoid nonspecific template interactions and eventually effectively avoid undesirable background amplification; the colorimetric assay is based on the SPEXPAR amplicons induced gold nanoparticles (AuNPs) color change; and the LFA detection principle involves the amplicons hybridization with designed probes. Under the optimized conditions, the colorimetric assay could detect miRNA-155 as low as 0.41 pM in less than 35 min; and LFA could detect as low as 100 pM in less than 40 min. Furthermore, the method was adopted to detect miRNA-155 in human serum samples with satisfactory recoveries. In summary, this study offered a promising analytical tool for molecular diagnostic testing application.
研究领域:
生物技术
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医学诊断
·
分子生物学
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产品关联
DOI
高影响因子
ACS Sensors
·
2025
·
IF 9.1
Jing Yang,Xiaoye Feng,Haiqin Li,Pengtao Chang,Xiaojun Ren,Xiaochun Li
Chronic kidney disease (CKD) has become a public health burden, with a high mortality rate and a trend toward youthfulness. It is imperative to develop a home monitoring system for the early diagnosis and long-term monitoring of CKD serum biomarkers. However, due to the limitations of conventional centrifugation preprocessing of whole blood samples, the real-time detection platform cannot fully meet the requirement of obtaining quantitative results directly from whole blood. Herein, a novel smartphone-assisted microfluidic chip integration of a whole blood separation point-of-care testing (POCT) system was proposed to achieve accurate quantification of CKD biomarkers uric acid (UA), creatinine (CR), and albumin (Alb) based on 30 μL of whole blood (fingertip blood). The whole blood separation module allows rapid separation of plasma from whole blood samples by vertical flow. A smartphone App for automated image recognition and processing was developed, which was applied for at-home routine healthcare. The detection limits of UA, CR, and Alb were 0.1127 mmol/L, 0.2978 μmol/L, and 0.7696 mg/mL, respectively, meeting clinical requirements. Additionally, the quantitative performance of 83 clinical whole blood samples showed high consistency with the clinical analyzer (Beckman AU 5400 automatic biochemical analyzer). This system is simple to operate and user-friendly, providing an accurate, rapid, cost-effective, and reliable detection method for the early screening and real-time monitoring of CKD, which holds significant potential in enhancing home self-management of CKD.
研究领域:
生物医学工程
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临床诊断
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微流控技术
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即时检测
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慢性肾病
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生物传感器
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家庭医疗健康技术
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL SCIENCES
·
2021
·Vol. 37
·期 12
·1695-1700
·
IF 2.081
Nailong GAO,Jianguo CHANG,Peng DAI,Ziming ZHU,Hui YOU
Alpha-fetoprotein (AFP) is an important disease biomarker, relating to cancers such as hepatocarcinomas and gastric cancer. However, traditional methods are time-consuming, relied on bulky instruments and trained professionals, cannot satisfy the demand for low cost and point-of-care testing (POCT). In this study, a power-free POCT device was developed for the rapid and low-cost detection of AFP via one-sampling. Based on the principle of sandwich immunofluorescence, the chip is capable of automatically accomplishing on-chip mixing, labeling and capturing procedures, which only require that operator add 40 μL sample into the chip one time. The proposed device is capable of sensitively detecting human AFP in FBS with a dynamic range of 10 – 1000 ng/mL and LOD (1.88 ng/mL) within a short time of 3 min. Predictably, our method holds a great potential to be applied in the POC diagnostics of proteins, especially for some regions that are resource-limited.
研究领域:
生物医学工程
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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
Biosensors-Basel
·
2022
·Vol. 12
·期 9
·712
·
IF 5.743
Qin Zhu,Huimin Wu,Zhen Ma,Yuqiao Liu,Junmin Li,Ling Zhu,Xinran Zhang,Chengcheng Wang,Dajing Chen,Danhua Zhu
In this paper, we report a point-of-care (POCT) testing strip based on a porous membrane structure for whole blood separation and colorimetric analysis without external supporting equipment. Conventional blood tests rely on large instruments for blood pretreatment and separation to improve measurement accuracy. Cellulose acetate (CA) membranes with different pore diameters and structures were prepared via a non-solvent method for the separation of whole blood. Among them,[email protected]membranes with nano-pores on the surface and micro-pores in the interior facilitated the capture of blood cells on the surface, as well as the free diffusion of plasma through the porous interior structure. The fluid flow of blood in the asymmetric porous structure can be theoretically estimated using the Lucas-Washburn equation. Compared with the conventional paper strips and other porous membranes, the[email protected]membrane with an immobilized sensing layer exhibited efficient blood separation, a short response time (less than 2 min), an ultralow dosage volume (5 μL), and high sensitivity. The fabricated blood separation membranes can be further used for the detection of various biomarkers in whole blood, providing additional options for rapid quantitative POCT tests.Keywords:blood separation;porous membranes;colorimetric assay
研究领域:
生物医学工程
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分析化学
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材料科学
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即时诊断
✓ 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）