point-of-care test

即时测试
  • 文章类型: Journal Article
    本文报道了一种快速灵敏的传感器,用于通过电化学机理检测和定量COVID-19N蛋白(N-PROT)。单频电化学阻抗谱用作基于金缀合物修饰的碳网印刷电极(Cov-Ag-SPE)的免疫传感器系统中实时测量N-PROT的转导方法。该系统通过在300s内的100Hz下由0.0VDC电势和10mVRMS-1AC信号组成的最佳刺激信号实现了高选择性。Cov-Ag-SPE在1.0ngmL-1至10.0μgmL-1的浓度下对N-PROT检测显示出对数响应,相位(θ)变化的相关系数为0.977。可以使用从阳性和阴性样本中观察到的某些方面创建基于ML的方法;因此,可以对252个样本进行分类,灵敏度达到83.0、96.2和91.3%,特异性,和准确性,分别,置信区间(CI)为73.0至100.0%。因为阻抗谱测量可以用低成本的便携式仪器进行,这里提出的免疫传感器可以应用于大规模检测的即时诊断,即使在资源有限的地方,作为常见诊断方法的替代方案。
    This paper reports a rapid and sensitive sensor for the detection and quantification of the COVID-19 N-protein (N-PROT) via an electrochemical mechanism. Single-frequency electrochemical impedance spectroscopy was used as a transduction method for real-time measurement of the N-PROT in an immunosensor system based on gold-conjugate-modified carbon screen-printed electrodes (Cov-Ag-SPE). The system presents high selectivity attained through an optimal stimulation signal composed of a 0.0 V DC potential and 10 mV RMS-1 AC signal at 100 Hz over 300 s. The Cov-Ag-SPE showed a log response toward N-PROT detection at concentrations from 1.0 ng mL-1 to 10.0 μg mL-1, with a 0.977 correlation coefficient for the phase (θ) variation. An ML-based approach could be created using some aspects observed from the positive and negative samples; hence, it was possible to classify 252 samples, reaching 83.0, 96.2 and 91.3% sensitivity, specificity, and accuracy, respectively, with confidence intervals (CI) ranging from 73.0 to 100.0%. Because impedance spectroscopy measurements can be performed with low-cost portable instruments, the immunosensor proposed here can be applied in point-of-care diagnostics for mass testing, even in places with limited resources, as an alternative to the common diagnostics methods.
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  • 文章类型: Journal Article
    弓形虫病,虽然通常无症状且作为食源性疾病流行,对怀孕期间免疫功能低下的个体造成相当大的死亡风险。在有限的资源下检测患者血清中特异性IgG和IgM的即时血清学测试对于疾病管理至关重要。尽管在商业试剂盒中用重组抗原(rAgs)替换弓形虫总裂解物抗原(TLA)的许多努力,而IgG检测提供了显着的特异性和灵敏度,IgM检测的灵敏度仍然相对较低。在这项研究中,我们试图在早期感染中鉴定靶向IgM的新型抗原,从而建立IgM现场检测试剂盒。用双向凝胶电泳(2DE)和小鼠血清免疫印迹,三种新型抗原,包括EF1γ,PGKI,和GAP50,用于靶向弓形虫IgM。然而,在免疫印迹验证实验中,小鼠血清IgM检测不到rAgEF1γ,用PGKI包被的ELISA没有消除交叉反应性,与GAP50相比。随后,使用涂有0.3mg/mL纯化rAgGAP50的条带的侧流反应,与基于速殖子TLA的常规ELISA相比,具有显着的灵敏度,它成功地鉴定了感染了速殖子的小鼠血清中的IgM,在5dpi时从103到104,在7dpi时从104,分别。此外,通过使用世界卫生组织的标准弓形虫感染的人血清,使用GAP50的快速荧光免疫色谱检测(FICT)的检测限(LOD)为0.65IU(国际单位).这些发现强调了GAP50的特定免疫反应性,表明其作为特异性生物标志物的潜力,可提高FCT在IgM检测中的敏感性。
    Toxoplasmosis, while often asymptomatic and prevalent as a foodborne disease, poses a considerable mortality risk for immunocompromised individuals during pregnancy. Point-of-care serological tests that detect specific IgG and IgM in patient sera are critical for disease management under limited resources. Despite many efforts to replace the T. gondii total lysate antigens (TLAs) by recombinant antigens (rAgs) in commercial kits, while IgG detection provides significant specificity and sensitivity, IgM detection remains comparatively low in sensitivity. In this study, we attempted to identify novel antigens targeting IgM in early infection, thereby establishing an IgM on-site detection kit. Using two-dimensional gel electrophoresis (2DE) and mouse serum immunoblotting, three novel antigens, including EF1γ, PGKI, and GAP50, were indicated to target T. gondii IgM. However, rAg EF1γ was undetectable by IgM of mice sera in Western blotting verification experiments, and ELISA coated with PGKI did not eliminate cross-reactivity, in contrast to GAP50. Subsequently, the lateral flow reaction employing a strip coated with 0.3 mg/mL purified rAg GAP50 and exhibited remarkable sensitivity compared with the conventional ELISA based on tachyzoite TLA, which successfully identified IgM in mouse sera infected with tachyzoites, ranging from 103 to 104 at 5 dpi and 104 at 7 dpi, respectively. Furthermore, by using standard T. gondii-infected human sera from WHO, the limit of detection (LOD) for the rapid fluorescence immunochromatographic test (FICT) using GAP50 was observed at 0.65 IU (international unit). These findings underline the particular immunoreactivity of GAP50, suggesting its potential as a specific biomarker for increasing the sensitivity of the FICT in IgM detection.
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  • 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
    自2019年底新型严重急性呼吸道综合征冠状病毒-2(SARS-CoV-2)爆发以来,该病毒的传播对公共卫生和全球经济构成了重大威胁。这项工作提出了一个步骤,双结构转换适体介导的信号放大级联,用于快速和灵敏地检测SARS-CoV-2核衣壳蛋白。该系统由两个具有结构转换功能的DNA适体和燃料DNA组成,其中链杂交和置换的级联触发了荧光产生和信号放大。这种基于适体的扩增级联既不需要使用酶的扩增阶段,也不需要预处理步骤如洗涤,病毒分离,和基因提取。该测定法可以将SARS-CoV-2与其他呼吸道病毒区分开,并在室温下30分钟内检测到高达1.0PFU/SARS-CoV-2的测定法。在35个鼻咽临床样本中,该测定法准确评估了25个阳性和10个阴性临床拭子样本,这是用定量聚合酶链反应证实的。本文报道的策略可以帮助检测液体样品中各种疾病的新出现的病原体和生物标志物。此外,开发的仅由DNA和荧光团组成的检测系统可以广泛集成到液体活检平台中以进行疾病诊断。
    Since the outbreak of the novel severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) at the end of 2019, the spread of the virus has posed a significant threat to public health and the global economy. This work proposed a one-step, dual-structure-switching aptamer-mediated signal amplification cascade for rapid and sensitive detection of the SARS-CoV-2 nucleocapsid protein. This system consisted of two DNA aptamers with structure-switching functionality and fuel DNA, where a cascade of strand hybridization and displacement triggered fluorescence generation and signal amplification. This aptamer-based amplification cascade required neither an amplification stage using enzymes nor pre-processing steps such as washing, viral isolation, and gene extraction. The assay could distinguish SARS-CoV-2 from other respiratory viruses and detect up to 1.0 PFU/assay of SARS-CoV-2 within 30 min at room temperature. In 35 nasopharyngeal clinical samples, the assay accurately assessed 25 positive and 10 negative clinical swab samples, which were confirmed using quantitative polymerase chain reaction. The strategy reported herein can help detect newly emerging pathogens and biomarkers of various diseases in liquid samples. In addition, the developed detection system consisting of only DNA and fluorophores can be widely integrated into liquid biopsy platforms for disease diagnosis.
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  • 文章类型: Journal Article
    这项研究提出了一种有效的基于纸的分析装置(PAD),用于使用由二维Fe/Ce金属有机框架(FeCe-BTC)制成的创新双纳米酶定量各种糖。MOF表现出优异的双功能过氧化物酶-氧化酶活性,高效催化鲁米诺的化学发光(CL)反应。作为过氧化物酶样纳米酶,FeCe-BTC能促进过氧化氢(H2O2)分解为羟基自由基,然后氧化鲁米诺。此外,还发现,当与H2O2反应时,MOF会变成混合价MOF,并充当氧化酶纳米酶。这种活性是由MOF结构中生成的Ce4+离子直接氧化鲁米诺引起的。MOF直接在PAD上合成,并与特定的天然酶级联,快速,和用于测量不同糖的选择性CL传感器。手机也被用来记录光强度,然后将其与分析物浓度相关联。设计的PAD对葡萄糖表现出0.1-10mM的宽线性范围,果糖,和蔗糖,检出限为0.03、0.04和0.04mM,分别。在食品和生物样品中显示出令人满意的结果,回收率为95.8至102.4%,这使其成为用于食品控制和医疗目的的即时护理(POC)测试的有希望的候选人。
    This study presents a potent paper-based analytical device (PAD) for quantifying various sugars using an innovative bi-nanozyme made from a 2-dimensional Fe/Ce metal-organic framework (FeCe-BTC). The MOF showed excellent bifunctional peroxidase-oxidase activities, efficiently catalyzing luminol\'s chemiluminescence (CL) reaction. As a peroxidase-like nanozyme, FeCe-BTC could facilitate the dissociation of hydrogen peroxide (H2O2) into hydroxyl radicals, which then oxidize luminol. Additionally, it was also discovered that when reacting with H2O2, the MOF turns into a mixed-valence MOF, and acts as an oxidase nanozyme. This activity is caused by the generated Ce4+ ions in the structure of MOF that can directly oxidize luminol. The MOF was directly synthesized on the PAD and cascaded with specific natural enzymes to establish simple, rapid, and selective CL sensors for the measurement of different sugars. A cell phone was also used to record light intensities, which were then correlated to the analyte concentration. The designed PAD showed a wide linear range of 0.1-10 mM for glucose, fructose, and sucrose, with detection limits of 0.03, 0.04, and 0.04 mM, respectively. It showed satisfactory results in food and biological samples with recovery values ranging from 95.8 to 102.4 %, which makes it a promising candidate for point-of-care (POC) testing for food control and medicinal purposes.
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  • 文章类型: Journal Article
    在资源丰富的地区每天使用即时测试(POCT),然而,在全球南部是有限的,特别是在院前设置。关于社区卫生工作者(CHW)使用非疟疾性POCT的研究很少。此范围审查的目的是描述院前环境中评估的POCT的当前多样性和广度。
    由经验丰富的医学图书馆员对已知关键文章进行医学主题标题(MeSH)分析,并在每个数据库中进行范围搜索,以捕获“护理点测试”和“社区卫生工作者”。“这项审查是由PRISMA扩展进行范围审查的指导。
    返回了2735个出版物,185人被提名进行全文审查,110项研究被证实符合研究标准.大多数人关注疟疾(74/110;67%)或HIV(25/110;23%);9/110(8%)描述了其他测试。这篇综述的结果表明,地理范围很广,当地CHW的术语具有显著的异质性。
    新POCT的使用正在增加,并且可能会在有限的资源环境中改善早期风险分层。来自数十年疟疾POCT的当前证据可以指导未来的实施战略。
    Point-of-Care Tests (POCTs) are utilized daily in resource abundant regions, however, are limited in the global south, particularly in the prehospital setting. Few studies exist on the use of non-malarial POCTs by Community Health Workers (CHWs). The purpose of this scoping review is to delineate the current diversity in and breadth of POCTs evaluated in the prehospital setting.
    A medical subject heading (MeSH) analysis of known key articles was done by an experienced medical librarian and scoping searches were performed in each database to capture \"point of care testing\" and \"community health workers.\" This review was guided by the PRISMA Extension for scoping reviews.
    2735 publications were returned, 185 were nominated for full-text review, and 110 studies were confirmed to meet study criteria. Majority focused on malaria (74/110; 67%) or HIV (25/110; 23%); 9/110 (8%) described other tests administered. Results from this review demonstrate a broad geographic range with significant heterogeneity in terminology for local CHWs.
    The use of new POCTs is on the rise and may improve early risk stratification in limited resource settings. Current evidence from decades of malaria POCTs can guide future implementation strategies.
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  • 文章类型: Journal Article
    背景:最近建议将低剂量氨甲环酸(TXA)用于体外循环(CPB),以减少相关并发症。尽管无法进行TXA浓度的即时实验室测试,在ClotPro®系统上进行的新型TPA测试可以测量TXA诱导的纤维蛋白溶解抑制。我们评估了TPA测试在体外和需要CPB的手术患者中的性能。
    方法:从六名志愿者中获取血液样本,用于体外评估组织纤溶酶原激活物(tPA)触发的纤维蛋白溶解和TXA的作用。随后在20名心脏手术患者中进行了一项观察性研究,以评估TXA对TPA测试的临床影响。
    结果:TXA≥2mgL-1以浓度依赖的方式抑制tPA诱导的纤溶亢进,并在TXA≥10mgL-1时完全抑制。在接受CPB的患者中,在CPB结束时0.1g推注TXA后,对TPA测试参数可检测到抗纤维蛋白溶解作用,TXA≥0.5g时,纤溶完全抑制。1gTXA对TPA测试参数的抗纤溶作用在术后18小时内逐渐减弱。然而,在估计肾小球滤过率(eGFR)≤30mlmin-11.73m-2的4例患者中,纤溶抑制持续.eGFR与术后18h的TPA检测参数有很强的相关性(r=0.86-0.92;P<0.0001)。
    结论:TPA试验对低浓度的TXA敏感,可作为心脏手术患者术后纤溶活性的实用监测工具。该测试可能对严重肾功能损害的患者特别有用。
    BACKGROUND: Low-dose tranexamic acid (TXA) has been recently recommended for cardiopulmonary bypass (CPB) to reduce associated complications. Although point-of-care laboratory tests for TXA concentrations are unavailable, a novel TPA-test on the ClotPro® system can measure TXA-induced inhibition of fibrinolysis. We evaluated the performance of the TPA-test in vitro and in patients undergoing surgery requiring CPB.
    METHODS: Blood samples were obtained from six volunteers for in vitro evaluation of tissue plasminogen activator (tPA)-triggered fibrinolysis and the effects of TXA. This was followed by an observational study in 20 cardiac surgery patients to assess clinical effects of TXA on the TPA-test.
    RESULTS: Hyperfibrinolysis induced by tPA was inhibited by TXA ≥2 mg L-1 in a concentration-dependent manner, and was completely inhibited at TXA ≥10 mg L-1. In patients undergoing CPB, antifibrinolytic effect was detectable on TPA-test parameters after a 0.1 g bolus of TXA at the end of CPB, and complete inhibition of fibrinolysis was obtained with TXA ≥0.5 g. The antifibrinolytic effects of 1 g TXA on TPA-test parameters were gradually attenuated over 18 h after surgery. However, the fibrinolytic inhibition continued in four patients with estimated glomerular filtration rate (eGFR) ≤30 ml min-1 1.73 m-2. The eGFR had strong correlations with TPA-test parameters at 18 h after surgery (r=0.86-0.92; P<0.0001).
    CONCLUSIONS: The TPA-test is sensitive to low concentrations of TXA and serves as a practical monitoring tool for postoperative fibrinolytic activity in cardiac surgery patients. This test might be particularly useful in patients with severe renal impairment.
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  • 文章类型: Journal Article
    猫白血病病毒(FeLV)是家猫的猫白血病综合征的原因。由FeLV引起的疾病的预防和控制主要基于疫苗接种以及感染受试者的识别和隔离。抗原诊断方法,在临床实践中使用最广泛的,可以与分子测试相关联以表征检测到的FeLV。在这项研究中,使用定量SYBRGreenReal-TimePCR(qPCR)检测2018-2021年转诊至意大利北部兽医教学医院的抗原阳性猫血液样本中的FeLV前病毒DNA.为了对鉴定出的病毒进行基因表征,使用6个不同的终点PCR扩增病毒包膜(env)基因的一部分并进行测序.研究中包括的26只猫中的22只(84.6%)通过qPCR测定测试为阳性。这表明所采用的qPCR的高性能,但需要进一步的研究来调查四只猫的抗原测试和qPCR之间不一致结果的原因。从env基因分析,15/22qPCR阳性猫被FeLV亚型A感染,5/15与B亚型同时感染。
    Feline leukemia virus (FeLV) is responsible for feline leukemia syndrome in domestic cats. The prevention and control of disease caused by FeLV are primarily based on vaccination and identification and isolation of infected subjects. Antigen diagnostic methods, which are the most widely used in clinical practices, can be associated to molecular tests to characterize the FeLV detected. In this study, a quantitative SYBR Green Real-Time PCR (qPCR) assay was used to detect FeLV proviral DNA in blood samples from antigen positive cats referred to a veterinary teaching hospital in Northern Italy in 2018-2021. To genetically characterize the identified viruses, a portion of the viral envelope (env) gene was amplified using six different end-point PCRs and sequenced. Twenty-two of 26 (84.6%) cats included in the study tested positive by qPCR assay. This suggests a high performance of the qPCR adopted but further studies are required to investigate the cause of discordant results between the antigen test and qPCR in four cats. From env gene analysis, 15/22 qPCR-positive cats were infected by FeLV subtype A and 5/15 shown coinfection with subtype B.
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  • 文章类型: Journal Article
    印度需要进行广泛的检测,以阻止新型冠状病毒在人群中的传播。虽然RT-PCR是推荐的诊断技术,由于需要专门的仪器,它的使用仅限于设备齐全的实验室,试剂和训练有素的人员。WHO尚未推荐用于诊断活动性感染的免疫诊断测试。非常需要开发即时分子测试。根据我们过去使用paperfluid设备通过分子测试诊断细菌感染的经验,我们建议开发COVID-19的诊断测试。作为一种平台技术,它可以适应未来的其他病毒爆发。
    There is a need for widespread testing in India to stop the spread of the novel coronavirus in the population. While RT-PCR is the recommended diagnostic technique, its use is limited to well-equipped laboratories due to the need for specialized instrumentation, reagents and trained personnel. Immunodiagnostic tests are not yet recommended by the WHO for diagnosing active infections. There is a strong need for developing point-of-care molecular tests. Based on our past experience with paperfluidic devices for diagnosing bacterial infections by molecular tests, we propose the development of a diagnostic test for COVID-19. As a platform technology, it could be adapted to other viral outbreaks in future.
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