point-of-care test

即时测试
  • 文章类型: Journal Article
    Antibiotics are widely utilized in agriculture for the prevention and treatment of animal diseases. How-ever, the abuse and overuse of antibiotics progressively increase the risks of antibiotic residues and antibiotic resis-tance. The bioaccumulation and biomagnification of antibiotics through food chains will negatively affect ecological safety, and finally threaten human health. There are many shortages of traditional antibiotic detection techniques, such as complex procedures, complicated operation and time consuming, and thus are difficult to meet the demand of instant, efficient and accurate on-site detection. Therefore, it is crucial to develop rapid detection techniques of antibiotics to manage the application of antibiotics in agriculture. We reviewed the utilization, and management of antibiotics in animal husbandry, residual characteristics, and potential hazards of antibiotics in agricultural products, summarized the advancements in rapid detection techniques of antibiotics in agricultural products over the past five years, compared the advantages and disadvantages of different rapid detection techniques, and prospected the future development in this area. This review would provide a valuable reference to the control and point-of-care test of antibiotics in agricultural products.
    在农业上,抗生素广泛用于动物的疾病预防和治疗。但抗生素的过度使用,甚至滥用,使得抗生素残留和抗生素耐药性问题日渐严重。抗生素通过食物链的生物富集和放大,将影响生态环境安全,并最终危害人体健康。传统的抗生素检测技术存在程序繁琐、操作复杂、耗时长等一系列问题,难以满足即时、高效和准确的现场检测需求。因此,为应对抗生素引起的食品安全问题、规范抗生素在农业上的使用,建立农产品抗生素的快速检测技术显得十分重要。本文综述了世界主要国家地区抗生素在养殖业的使用和管理情况,以及抗生素在农产品中的残留特征和对生物体及环境的危害,归纳了近5年内农产品中抗生素快速检测技术的发展情况,对各项快速检测技术的优缺点进行了对比,最后对未来发展方向进行了展望。本文可为农产品抗生素管控和即时检测提供借鉴。.
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  • 文章类型: Journal Article
    尽管纳米粒子镜腔的灵敏度很高,等离子体纳米材料的一个重要分支,复杂的制备和读出过程限制了其在生物传感中的广泛应用。或者,液态金属(LMs)结合了流动性和出色的等离子体特性,已成为构建等离子体纳米结构的潜在候选者。在这里,我们提出了一种微流控集成策略来构建基于LM的免疫分析平台,使基于LM的纳米等离子体传感器可用于即时护理(POC)临床生物标志物检测。将可流动的LM引入到蛋白质涂覆的Au纳米颗粒单层上,形成“镜面纳米颗粒”纳米结构,简化了在常规的纳米颗粒镜腔中的制造工艺。当抗体被包被在Au纳米颗粒单层上的抗原捕获时,器件响应生物分子层的厚度和折射率变化,输出肉眼可读信号,具有高灵敏度(检测限:~604fM)和宽动态范围(6阶)。这种新的检测方法,在30分钟内产生定量结果,允许高吞吐量,基于智能手机的SARS-CoV-2抗体检测针对临床血清或血液样本中的多种变体。这些结果为使用LM材料进行POC测试建立了先进的途径,并证明其促进诊断的潜力,各种传染病的监测和流行研究。
    Despite high sensitivity of nanoparticle-on-mirror cavities, a crucial branch of plasmonic nanomaterials, complex preparation and readout processes limit their extensive application in biosensing. Alternatively, liquid metals (LMs) combining fluidity and excellent plasmonic characteristics have become potential candidates for constructing plasmonic nanostructures. Herein, we propose a microfluidic-integration strategy to construct LM-based immunoassay platform, enabling LM-based nanoplasmonic sensors to be used for point-of-care (POC) clinical biomarker detection. Flowable LM is introduced onto protein-coated Au nanoparticle monolayer to form a \"mirror-on-nanoparticle\" nanostructure, simplifying the fabrication process in the conventional nanoparticle-on-mirror cavities. When antibodies were captured by antigens coated on the Au nanoparticle monolayer, devices respond both thickness and refractive index change of biomolecular layers, outputting naked-eye readable signals with high sensitivity (limit of detection: ∼ 604 fM) and a broad dynamic range (6 orders). This new assay, which generates quantitative results in 30 min, allows for high-throughput, smartphone-based detection of SARS-CoV-2 antibodies against multiple variants in clinical serum or blood samples. These results establish an advanced avenue for POC testing with LM materials, and demonstrate its potential to facilitate diagnostics, surveillance and prevalence studies for various infectious diseases.
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  • 文章类型: Journal Article
    提出了通过用胶带从Cu片上剥离化学印刷的银纳米珊瑚(AgNCs)来制备夹心等离子体耦合表面增强拉曼光谱(SERS)胶带,它可以从食品表面取样目标,并将它们夹在基底和Cu片之间进行SERS检测。制备SERS胶带的固-固转化方法可以有效避免制备过程中胶带粘性的减弱。AgNC衬底的夹心等离子体耦合结构,目标,Cu片显示出优异的SERS活性(EF=1.62×107),可用于分析物的灵敏测定。此外,由于铜片的高导热性,SERS检测过程中激光辐照的热效应不会损坏AgNC带,这确保了后续定量的可重复性。通过在不同浓度范围下的两个典型校准图,证明了夹心等离子体耦合SERS带可以以良好的线性系数(>0.98)量化孔雀石绿(MG)和甲基对硫磷(MP)。对于MG和MP,该方法的检出限(LOD)为0.17ng/cm2和0.48μg/cm2(S/N=3)。该方法可以实现水果和鱼鳞表面MP和MG的定量测定,回收率为93-113%。令人满意的检测结果表明,所提出的夹心等离子体耦合AgNC胶带可以成功地应用于基于SERS的即时检测(POCT),以测定农药残留。这将为设计和构造SERS磁带提供新的途径。
    A sandwich plasmonic coupled surface enhanced Raman spectroscopy (SERS) tape is proposed prepared by peeling the chemical printed silver nanocorals (AgNCs) from Cu sheet with adhesive tape, which can sample targets from food surface and sandwich them between substrates and Cu sheet for SERS detection. The solid-to-solid transformation method for fabricating SERS tapes can effectively avoid the weakening of tape stickiness during the preparation process. The sandwich plasmonic coupled structure of AgNC substrate, targets, and Cu sheet display excellent SERS activity (EF = 1.62 × 107) for sensitive determination of analytes. In addition, due to the high heat conductivity of Cu sheet, the thermal effect of laser irradiation during SERS detection cannot damage the AgNC tapes, which ensures the reproducibility of subsequent quantification. The sandwich plasmonic coupled SERS tape is demonstrated to quantify malachite green (MG) and methyl parathion (MP) with good linear coefficients (> 0.98) by two typical calibration plots under different concentration ranges. The limit of detection (LOD) of the method is 0.17 ng/cm2 and 0.48 μg/cm2 (S/N = 3) for MG and MP. This method can realize the quantitative determination of MP and MG on the surface of fruits and fish scale with recoveries of 93-113%. The satisfactory detection results demonstrate the proposed sandwich plasmonic coupled AgNC tape can be successfully applied to SERS-based point-of-care testing (POCT) for pesticide residue determination, which will provide a new path for designing and constructing SERS tapes.
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  • 文章类型: Journal Article
    宫颈癌是全球威胁女性健康的第三大常见癌症,高危型人乳头瘤病毒(HR-HPV)感染是全球宫颈癌的主要病因。鉴于HR-HPV感染的复发性,准确的筛选对其控制至关重要。由于常用的聚合酶链反应(PCR)技术受到专业设备和人员的限制,仍然非常需要方便和超灵敏的HR-HPV检测方法.作为新的分子检测方法,基于核酸扩增的生物传感器具有灵敏度高的优点,快速操作,和便携性,这有助于农村和偏远地区的即时检测。本文综述了基于多种核酸扩增策略在改良PCR中用于HR-HPV筛查的核酸生物传感器。环介导等温扩增,重组酶聚合酶扩增,杂交链反应,催化发夹组件,和CRISPR/Cas系统。结合微流控技术,侧流测定,电化学分析和其他传感技术,HR-HPV核酸生物传感器具有高通量,响应时间短,高灵敏度和易于操作的领域。虽然还有不足之处,如成本高,重现性差,这种方法将适用于HR-HPV感染或宫颈癌的现场筛查,以及未来在复杂环境和贫困地区的辅助临床诊断.
    Cervical cancer is the third most common cancer threatening women\'s health globally, and high-risk human papillomavirus (HR-HPV) infection is the main cause of cervical cancer worldwide. Given the recurrent nature of HR-HPV infection, accurate screening is essential for its control. Since the commonly used polymerase chain reaction (PCR) technique is limited by professional equipment and personnel, convenient and ultrasensitive detection methods for HR-HPV are still highly needed. As new molecular detection methods, nucleic acid amplification-based biosensors have the advantages of high sensitivity, rapid operation, and portability, which are helpful for point-of-care testing in rural and remote areas. This review summarized nucleic acid biosensors for HR-HPV screening based on a variety of nucleic acid amplification strategies involved in improved PCR, loop-mediated isothermal amplification, recombinase polymerase amplification, hybridization chain reaction, catalyzed hairpin assembly, and CRISPR/Cas systems. In combination with microfluidic technology, lateral flow assays, electrochemical analysis and other sensing technologies, HR-HPV nucleic acid biosensors have the advantages of high throughput, short response time, high sensitivity and easy operation in the field. Although there are still shortcomings, such as high cost and poor reproducibility, this approach will be suitable for on-site screening of HR-HPV infection or cervical cancer and for auxiliary clinical diagnosis in complex environments and poor areas in the future.
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  • 文章类型: Journal Article
    活的食源性病原体,具有快速增殖能力和高致病性,食品安全和公共卫生危机。病原体的高灵敏度检测尤其必要,但仍然具有挑战性。这项工作开发了一种功能化的尼龙拭子阵列,该阵列对鼠伤寒沙门氏菌(S.T.)具有增强的亲和力,用于基于高特异性ATP生物发光的S.T.检测。简而言之,尼龙拭子(NyS)通过与甲醛反应转变为N-甲基化尼龙(NyS-OH),和NyS-OH通过与柠檬酸(CA)和EDC/NHS溶液的羧基反应进一步转化为NyS-CA,用于改变NyS表面能以有利于生物改性。抗体固定的尼龙拭子(MNyS-Ab)准备用于S.T.特异性吸附。将三个制备的MNyS-Ab安装在搅拌器上以形成MNyS-Ab阵列,允许现场富集S.T.通过吸收提取。通过使用便携式ATP生物发光传感器测量从细胞裂解释放的ATP的生物发光来定量富集的S.T.。生物测定法证实了在35分钟内的可检测范围为102-107CFUmL-1,检测限(LOD)为8CFU/mL。单个MNyS-Ab拭子的信号比直接检测106CFU/mLS.T强500倍。与单个MNyS相比,MNyS-Ab阵列显示提取水平增加100倍。便携式生物发光传感器和改性尼龙拭子阵列的这种组合为现场护理测试S.T.菌株提供了一种新颖的策略。它有望对其他病原体和病毒进行高灵敏度测量。
    Live food-borne pathogens, featured with rapid proliferative capacity and high pathogenicity, pose an emerging food safety and public health crisis. The high-sensitivity detection of pathogens is particularly imperative yet remains challenging. This work developed a functionalized nylon swab array with enhanced affinity for Salmonella typhimurium (S.T.) for high-specificity ATP bioluminescence-based S.T. detection. In brief, the nylon swabs (NyS) were turned to N-methylation nylon (NyS-OH) by reacting with formaldehyde, and NyS-OH were further converted to NyS-CA by reacting with carboxylic groups of citric acid (CA) and EDC/NHS solution, for altering the NyS surface energy to favor biomodification. The antibody-immobilized nylon swab (MNyS-Ab) was ready for S.T.-specific adsorption. Three prepared MNyS-Ab were installed on a stirrer to form an MNyS-Ab array, allowing for on-site enrichment of S.T. through absorptive extraction. The enriched S.T. was quantified by measuring the bioluminescence of ATP released from cell lysis utilizing a portable ATP bioluminescence sensor. The bioassay demonstrated a detectable range of 102-107 CFU mL-1 with a detection limit (LOD) of 8 CFU/mL within 35 min. The signal of single MNyS-Ab swabs was 500 times stronger than the direct detection of 106 CFU/mL S.T. The MNyS-Ab array exhibited a 100-fold increase in extraction level compared to a single MNyS. This combination of a portable bioluminescent sensor and modified nylon swab array offers a novel strategy for point-of-care testing of live S.T. strains. It holds promise for high-sensitivity measurements of other pathogens and viruses.
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  • 文章类型: Journal Article
    背景:葡萄糖和胆固醇的准确检测在疾病诊断和家庭护理中起着关键作用。为此,生化分析仪已成为广泛用于测量疾病生物标志物的工具。尽管如此,它们的便携性差和高成本限制了它们的可访问性,将它们的使用限制在实验室环境中,并阻碍了即时检测(POCT)的采用。相比之下,便携式和负担得起的纸质测试平台的出现通过提供距离信号彻底改变了诊断测试,增强直观性和视觉可达性。因此,这些平台越来越适合POCT。
    结果:我们开发了一个POCT平台,该平台集成了AuNS@Ag,刺激响应水凝胶和试纸,使葡萄糖的视觉距离读取。将银涂覆的AuNS和酶封装在温度响应性N-异丙基丙烯酰胺-丙烯酰胺(NIPAM-AcAm)水凝胶中,分别用作目标识别和反应单元。葡萄糖可以在水凝胶多孔基质中自由扩散,从而引发酶催化的反应,从而引起系统光热效应的改变。该动态过程确保系统的光热特性的有效和响应调制。通过巧妙地捕获由光热效应介导的水释放引起的距离信号,实现了目标物质的可视化和定量。线性范围从0到30mM。基于距离的POCT平台与商用血糖仪的一致性表明,该平台提供了一种便携式、经济实惠和可靠的方法视觉读取生物标志物。
    结论:提出的策略可以直接,目标的可视化定量分析,而不需要额外的分析仪器。特别是,该方法作为胆固醇和其他疾病标志物测量的有效平台具有重要前景.
    BACKGROUND: The accurate detection of glucose and cholesterol plays a pivotal role in disease diagnosis and home care. To this end, biochemical analyzers have become extensively utilized tools for measuring disease biomarkers. Nonetheless, their poor portability and high cost have restricted their accessibility, limiting their use to laboratory settings and hindering the adoption of point-of-care testing (POCT). In contrast, the emergence of portable and affordable paper-based testing platform has revolutionized diagnostic testing by providing distance signals, enhancing intuitiveness and visual accessibility. Consequently, these platforms have become increasingly suitable for POCT.
    RESULTS: We have developed a POCT platform that integrated AuNS@Ag, stimulus-responsive hydrogel and test strips, enabling visual distance reading of glucose. The silver-coated AuNS and enzyme were encapsulated within a temperature-responsive N-isopropylacrylamide-acrylamide (NIPAM-AcAm) hydrogel to act as target recognition and reaction units respectively. Glucose can diffuse freely within the hydrogel porous matrix, thereby instigating enzyme-catalyzed reaction that induce alterations in the photothermal effect of the system. This dynamic process ensures efficient and responsive modulation of the system\'s photothermal properties. By ingeniously capturing distance signals induced by the photothermal effect-mediated water release the visualization and quantification of target substances are achieved, with a linear range spanning from 0 to 30 mM. The consistency between distance-based POCT platform and commercial blood glucose meter demonstrates that the platform provides a portable, affordable and reliable method for visual reading biomarkers.
    CONCLUSIONS: The proposed strategy enables direct, visual quantitative analysis of the target without the need for additional analytical instruments. Particularly, this method holds significant promise as an efficient platform for cholesterol and other disease markers measurement.
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  • 文章类型: Journal Article
    新型高通量蛋白质检测技术对于基于人群的大规模SARS-CoV-2抗体检测以及监测针对病毒变体的免疫质量和持续时间是至关重要的。当前的蛋白质微阵列技术严重依赖于标记的转导方法,这些方法需要复杂的仪器和复杂的操作,限制了他们的临床潜力,特别是对于即时护理(POC)应用。这里,我们开发了一种基于厚度感应等离子体标尺的无标签和肉眼可读的微阵列(NRM),在30分钟内实现抗体谱分析。NRM芯片通过有效筛选抗原类型和实验条件,为中和抗体检测提供100%的准确性,并允许在临床样品中针对多种SARS-CoV-2变体的抗体谱分析。我们进一步建立了一个灵活的“条形码”NRM测定与一个简单的基于磁带的操作,实现基于智能手机的有效读出和分析。这些结果证明了高通量蛋白质检测的新策略,并突出了新型蛋白质微阵列技术在实际临床应用中的潜力。
    Novel high-throughput protein detection technologies are critically needed for population-based large-scale SARS-CoV-2 antibody detection as well as for monitoring quality and duration of immunity against virus variants. Current protein microarray techniques rely heavily on labeled transduction methods that require sophisticated instruments and complex operations, limiting their clinical potential, particularly for point-of-care (POC) applications. Here, we developed a label-free and naked-eye readable microarray (NRM) based on a thickness-sensing plasmon ruler, enabling antibody profiling within 30 min. The NRM chips provide 100% accuracy for neutralizing antibody detection by efficiently screening antigen types and experimental conditions and allow for the profiling of antibodies against multiple SARS-CoV-2 variants in clinical samples. We further established a flexible \"barcode\" NRM assay with a simple tape-based operation, enabling an effective smartphone-based readout and analysis. These results demonstrate new strategies for high-throughput protein detection and highlight the potential of novel protein microarray techniques for realistic clinical applications.
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  • 文章类型: Journal Article
    这项研究旨在通过利用增强的廷德尔效应(TE)现象来开发一种可靠且定量的方法来测量肌酐水平。设想的传感测定设计用于资源有限的环境或家庭中的实际部署,先进的实验室设施有限。其主要目标是能够定期和方便地监测肾脏保健,特别是在涉及肌酐水平升高的病例中。肌酐传感策略是基于通过肌酐和AuNP之间的直接交联反应触发的金纳米颗粒(AuNP)的聚集来实现的。使用廉价的激光指示器作为手持光源,使用智能手机作为便携式设备,记录肌酐诱导的AuNPs聚集增强的TE现象.在对AuNP浓度和TE测量时间等参数进行评估和优化后,随后的概念验证实验表明,TE图像的平均灰度值变化与肌酐浓度在1-50μM范围内的对数线性相关,检测限为0.084μM。同时,我们提出的肌酐传感平台在复杂的基质环境中表现出高度选择性的检测.我们的方法提供了一个简单的,成本效益高,和便携式肌酐检测手段,提出了一种令人鼓舞的信号读出机制,适用于即时护理(POC)应用。利用该测定作为POC解决方案显示出加快及时干预和增强患有肾脏健康问题的个体的医疗保健结果的潜力。
    This research aims to develop a robust and quantitative method for measuring creatinine levels by harnessing the enhanced Tyndall effect (TE) phenomenon. The envisioned sensing assay is designed for practical deployment in resource-limited settings or homes, where access to advanced laboratory facilities is limited. Its primary objective is to enable regular and convenient monitoring of renal healthcare, particularly in cases involving elevated creatinine levels. The creatinine sensing strategy is achieved based on the aggregation of gold nanoparticles (AuNPs) triggered via the direct crosslinking reaction between creatinine and AuNPs, where an inexpensive laser pointer was used as a handheld light source and a smartphone as a portable device to record the TE phenomenon enhanced by the creatinine-induced aggregation of AuNPs. After evaluation and optimization of parameters such as AuNP concentrations and TE measurement time, the subsequent proof-of-concept experiments demonstrated that the average gray value change of TE images was linearly related to the logarithm of creatinine concentrations in the range of 1-50 μM, with a limit of detection of 0.084 μM. Meanwhile, our proposed creatinine sensing platform exhibited highly selective detection in complex matrix environments. Our approach offers a straightforward, cost-effective, and portable means of creatinine detection, presenting an encouraging signal readout mechanism suitable for point-of-care (POC) applications. The utilization of this assay as a POC solution exhibits potential for expediting timely interventions and enhancing healthcare outcomes among individuals with renal health issues.
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  • 文章类型: Journal Article
    迫切需要便携式,灵敏的即时测试(POCT)方法来检测严重的急性呼吸道综合症冠状病毒2(SARS-CoV-2),以确保公众健康和安全。然而,从环境中检测真实水样中的痕量病原体仍然面临挑战,因为复杂的环境破坏者可以迅速降级目标。在这里,引入了用岩藻依聚糖和聚多巴胺(Fuc-PDA-MBs)包被的磁珠作为样品预处理的捕获载体。褐藻多糖,硫酸多糖,可以识别SARS-CoV-2刺突(S1)蛋白受体结合域(S1RBD),并被选择用于替代富集中的抗体。用SARS-CoV-2刺突假病毒接种的环境水用于Fuc-PDA-MBs方法的性能测试。在最优条件下,在107.62至104.34基因拷贝(gc)/L的SARS-CoV-2刺突假病毒浓度下,Fuc-PDA-MB的使用显示平均76%的捕获效率。与电负性过滤(ENF)相比,Fuc-PDA-MB显示出更好的病毒体吸附效果。Fuc-PDA-MB还通过原始污染的城市废水进行了验证,并显示出SARS-CoV-2变体的高回收率。为了快速检测POCT中的病毒,无核酸提取环介导等温扩增(LAMP)用于简化实验过程。Fuc-PDA-MBs-LAMP分析显示样品的定量限(LOQ)为105.49gc/L。整个过程可以在90分钟内完成,包括病毒预富集30分钟,10分钟核酸释放和45分钟LAMP分析。与常规的基于抗体的免疫检测相比,这个集成系统提供了广谱,在复杂环境中检测SARS-CoV-2突变体的经济方法,也适用于高通量测试,可用于发展中地区SARS-CoV-2疫情的现场预警。
    The portable and sensitive point-of-care-test (POCT) method is in urgent need to detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) for ensuring public health and safety. However, detection of trace number of pathogens in real water sample from the environment still faces challenges, because complex environment disruptors can rapidly degrade targets. Herein, magnetic beads coated with fucoidan and polydopamine (Fuc-PDA-MBs) were introduced as the capture carrier for pretreatment of samples. Fucoidan, a sulfated polysaccharide, can recognize the SARS-CoV-2 spike (S1) protein receptor-binding domain (S1 RBD) and was chosen for replacement of antibody in enrichment. Environmental water seeded with SARS-CoV-2 spike pseudovirus was applied to test performance of Fuc-PDA-MBs method. Under optimal conditions, the use of Fuc-PDA-MBs showed average 76 % capture efficiency at SARS-CoV-2 spike pseudovirus concentration ranging from 107.62 to 104.34 gene copies (gc)/L. Compared with Electronegative filtration (ENF), Fuc-PDA-MBs showed better virion sorption effectiveness. Fuc-PDA-MBs also validated by raw contaminated urban wastewater and showed high recovery results for SARS-CoV-2 variants. To rapidly detect virus in POCT, nucleic acid extraction-free Loop-Mediated Isothermal Amplification (LAMP) was used for simplifying experimental process. The Fuc-PDA-MBs-LAMP assay showed the quantitation limit of sample (LOQ) was 105.49 gc/L. The whole procedure could be completed within 90 min, including 30 min for virus pre-enrichment, 10 min nucleic acid release and 45 min LAMP analysis. Compared with regular antibody-based immunodetection, this integrated system provides broad-spectrum, economic way to detect SARS-CoV-2 mutants in complex environments and also adaptable for high throughput test, which might be used for on-site early warning of SARS-CoV-2 outbreaks in developing area.
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  • 文章类型: Journal Article
    “测试和去”单核苷酸变异(SNV)在几分钟内的检测仍然具有挑战性,特别是在低丰度样本中,因为现有方法面临灵敏度和测试速度之间的权衡。灵敏的检测通常依赖于复杂且耗时的核酸扩增或测序。这里,我们开发了由Argonaute蛋白介导的石墨烯场效应晶体管(GFET)平台,敏感和特异性SNV检测。Argonaute蛋白提供了一个纳米级的结合通道来预先组织DNA探针,在未扩增的肿瘤相关microRNA中以单核苷酸分辨率加速靶结合并快速识别SNV,循环肿瘤DNA,当在种子区域发生错配时,病毒RNA和逆转录cDNA。集成的微芯片在5分钟内同时检测与测序结果一致的多个SNV,以“test-and-go”方式实现最快的SNV检测,无需核酸提取,逆转录和扩增。本文受版权保护。保留所有权利。
    \"Test-and-go\" single-nucleotide variation (SNV) detection within several minutes remains challenging, especially in low-abundance samples, since existing methods face a trade-off between sensitivity and testing speed. Sensitive detection usually relies on complex and time-consuming nucleic acid amplification or sequencing. Here, a graphene field-effect transistor (GFET) platform mediated by Argonaute protein that enables rapid, sensitive, and specific SNV detection is developed. The Argonaute protein provides a nanoscale binding channel to preorganize the DNA probe, accelerating target binding and rapidly recognizing SNVs with single-nucleotide resolution in unamplified tumor-associated microRNA, circulating tumor DNA, virus RNA, and reverse transcribed cDNA when a mismatch occurs in the seed region. An integrated microchip simultaneously detects multiple SNVs in agreement with sequencing results within 5 min, achieving the fastest SNV detection in a \"test-and-go\" manner without the requirement of nucleic acid extraction, reverse transcription, and amplification.
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