rapid

RAPID
  • 文章类型: Journal Article
    基于导波阵列的结构健康监测(SHM)是诊断金属连接结构损伤的一种有前途的解决方案。在这个领域,概率检查的重建算法(RAPID)是用于执行损伤定位的最广泛使用的算法之一。在本文中,提出了一种基于阵列补偿损伤指数的密度聚类RAPID。基于新的损伤指数构造了一个新的概率分布函数,适应传感器阵列中的不同元件以补偿性能变化。然后,对RAPID算法的成像矩阵进行密度聚类以获得损伤的位置和程度。最后,该方法在加筋铝板上进行了实验验证。实验结果表明,该方法实现了损伤定位,实现了损伤的定量诊断。
    Guided wave array-based structural health monitoring (SHM) is a promising solution for diagnosing damage in metal-connected structures. In this field, the reconstruction algorithm for probabilistic inspection (RAPID) is one of the most widely used algorithms for performing damage localization. In this paper, a density clustering RAPID based on an array-compensated damage index is proposed. A new probability distribution function was constructed based on a new damage index, which is adaptive to different elements in the sensor array to compensate for performance variation. Then, the imaging matrix of the RAPID algorithm was density-clustered to obtain the location and degree of damage. Finally, the method was verified by experiments on a stiffened aluminum plate. The experimental results demonstrate that the method achieves damage localization and enables quantitative damage diagnosis.
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  • 文章类型: Journal Article
    将肝移植扩展到新的肿瘤学适应症可能会加剧移植物的短缺。活体肝移植(LDLT)可能成为一种可行的资源,尽管它在西方世界的传播仍然非常有限。几个团体主张通过减少供体肝切除术的程度来最大程度地减少对供体的影响。即,从右叶转移到左叶或左侧段捐赠(“向左转移”)。这在处理未确定的适应症时尤其重要,并且可以使潜在捐助者和接受者都更容易接受。左移植物可以直接移植,尽管小号综合症的风险更高,或者它们可以用于双移植LDLT或RAPID程序的设置,尽管技术复杂。这篇综述将揭示每种技术最相关的功能,突出他们的优势和陷阱,关注结果。大量的移植中心应该可以使用这套广泛的工具,提出适应供体-受体匹配的最佳技术。
    The extension of liver transplantation to new oncologic indications might exacerbate the shortage of grafts. Living donor liver transplantation (LDLT) may emerge as a viable resource, although its diffusion in the Western world is still very limited. Several groups have advocated for minimizing the impact on donors by reducing the extent of donor hepatectomy, i.e., shifting from right-lobe to left-lobe or left-lateral segment donation (\"shift-to-left\"). This is particularly relevant when dealing with non-established indications and could make it more acceptable both for potential donors and for the recipients. Left grafts can be transplanted straightforward, despite a higher risk of small-for-size syndrome, or they can be used in the setting of dual-graft LDLT or RAPID procedures, despite technical complexity. This review will expose the most relevant features of each technique, highlighting their strengths and pitfalls and focusing on outcomes. This wide set of tools should be available at high-volume transplant centers, to propose the best technique to adapt to donor-recipient matching.
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  • 文章类型: Journal Article
    快速,简单,低成本的诊断技术是对抗传染病的重要工具。我们描述了一类基于适体的RNA开关或aptaswitches,其识别靶核酸分子并启动报告适体的折叠。Aptaswitches几乎可以检测任何序列,并提供强烈的荧光读数,而无需介入酶,在短短5分钟内产生信号,并以最少的设备通过眼睛进行检测。Aptaswitches可用于调节七个荧光适体的折叠,提供了控制适体的一般手段和可复用的报告颜色的阵列。等温扩增反应与aptaswitches偶联,在一锅反应中,我们的灵敏度低至1个RNA拷贝/μL。针对来自临床唾液样品的RNA的多重一体化反应的应用对于在30分钟内检测SARS-CoV-2产生96.67%的总体准确度。因此,Aptaswitches是用于核酸检测的通用工具,其易于整合到快速诊断测定中。
    Rapid, simple, and low-cost diagnostic technologies are crucial tools for combatting infectious disease. We describe a class of aptamer-based RNA switches or aptaswitches that recognize target nucleic acid molecules and initiate folding of a reporter aptamer. Aptaswitches can detect virtually any sequence and provide an intense fluorescent readout without intervening enzymes, generating signals in as little as 5 minutes and enabling detection by eye with minimal equipment. Aptaswitches can be used to regulate folding of seven fluorogenic aptamers, providing a general means of controlling aptamers and an array of multiplexable reporter colors. Coupling isothermal amplification reactions with aptaswitches, we reach sensitivities down to 1 RNA copy/μL in one-pot reactions. Application of multiplexed all-in-one reactions against RNA from clinical saliva samples yields an overall accuracy of 96.67% for detection of SARS-CoV-2 in 30 minutes. Aptaswitches are thus versatile tools for nucleic acid detection that are readily integrated into rapid diagnostic assays.
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  • 文章类型: Journal Article
    侵袭性真菌感染是一种主要的健康威胁,发病率和死亡率高。强调迫切需要快速诊断工具来检测抗真菌耐药性。传统的基于培养的抗真菌药敏试验(AFST)方法由于其漫长的过程而常常达不到。在我们之前的研究中,我们开发了一种全载玻片成像(WSI)技术,用于细菌抗生素耐药性的高通量评估.建立在这个基础上,这项研究通过高通量监测数百种单个真菌的生长,使其适应快速AFST,从而扩大了WSI的应用范围。由于真菌独特的“萌芽”生长模式,我们开发了一种独特的方法,利用特定的细胞数量变化来确定真菌的复制,而不是在我们之前的研究中用于细菌的细胞面积变化,以准确确定单个真菌细胞的生长速率。该方法不仅通过直接观察单个真菌细胞的生长来加速抗真菌耐药性的测定,但也产生准确的结果。采用白色念珠菌作为代表性的模式生物,可靠的最低抑制浓度(MIC)的氟康唑抑制白色念珠菌的100%细胞(表示为MIC100)在3h内使用开发的方法获得,而改良的肉汤稀释法需要72h才能获得类似的可靠结果。此外,我们的方法被有效地用于直接检测血液培养样本,消除了从掺有白色念珠菌的全血样品中分离真菌的需要。这些特征表明,所开发的方法具有巨大的潜力,可作为快速抗真菌药敏试验和MIC测定的通用工具。
    Invasive fungal infections are a major health threat with high morbidity and mortality, highlighting the urgent need for rapid diagnostic tools to detect antifungal resistance. Traditional culture-based antifungal susceptibility testing (AFST) methods often fall short due to their lengthy process. In our previous research, we developed a whole-slide imaging (WSI) technique for the high-throughput assessment of bacterial antibiotic resistance. Building on this foundation, this study expands the application of WSI by adapting it for rapid AFST through high-throughput monitoring of the growth of hundreds of individual fungi. Due to the distinct \"budding\" growth patterns of fungi, we developed a unique approach that utilizes specific cell number change to determine fungi replication, instead of cell area change used for bacteria in our previous study, to accurately determine the growth rates of individual fungal cells. This method not only accelerates the determination of antifungal resistance by directly observing individual fungal cell growth, but also yields accurate results. Employing Candida albicans as a representative model organism, reliable minimum inhibitory concentration (MIC) of fluconazole inhibiting 100% cells of Candida albicans (denoted as MIC100) was obtained within 3h using the developed method, while the modified broth dilution method required 72h for the similar reliable result. In addition, our approach was effectively utilized to test blood culture samples directly, eliminating the need to separate the fungi from whole blood samples spiked with Candida albicans. These features indicate the developed method holds great potential serving as a general tool in rapid antifungal susceptibility testing and MIC determination.
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  • 文章类型: Journal Article
    获得安全和营养的食物对于维持生命和支持健康至关重要。食用被病原体污染的食物会导致从腹泻到癌症的严重疾病。许多食源性感染可导致长期损害甚至死亡。因此,早期检测食源性致病菌如致病性大肠杆菌菌株对公共安全至关重要。检测这些细菌的常规方法基于在选择性培养基上培养并遵循标准生化鉴定。尽管他们的准确性,这些方法是耗时的。基于PCR的病原体检测依赖于复杂的设备和专业的技术人员,这些设备和技术人员在资源有限的地区很难找到。而CRISPR技术对于鉴别致病菌更具特异性和敏感性,因为它采用了针对特定DNA序列的可编程CRISPR-Cas系统,最小化非特异性结合和交叉反应性。在这个项目中,提出了一种基于CRISPR-Cas12a传感的鲁棒检测方法,这是快速的,对从田纳西州17个农场的成年山羊的粪便样品中收集的致病性大肠杆菌分离株具有敏感性和特异性。检测反应中含有致病区PCR扩增产物,记者探测,Cas12a酶,和对三个致病基因stx1,stx2和hlyA具有特异性的crRNA。当在UV光下激发时,与致病菌的CRISPR反应发射荧光。为了评估该测定法的检测灵敏度和特异性,将其结果与基于PCR的检测方法进行比较。两种方法对于相同的样品产生相似的结果。这项技术非常精确,高度敏感,快,成本有效,易于使用,并且可以容易地克服现有检测方法的局限性。该项目可以产生一种通用的检测方法,该方法易于适应快速检测和监测对人类健康构成大规模生物安全威胁的疾病,植物和动物生产。
    Access to safe and nutritious food is critical for maintaining life and supporting good health. Eating food that is contaminated with pathogens leads to serious diseases ranging from diarrhea to cancer. Many foodborne infections can cause long-term impairment or even death. Hence, early detection of foodborne pathogens such as pathogenic Escherichia coli strains is essential for public safety. Conventional methods for detecting these bacteria are based on culturing on selective media and following standard biochemical identification. Despite their accuracy, these methods are time-consuming. PCR-based detection of pathogens relies on sophisticated equipment and specialized technicians which are difficult to find in areas with limited resources. Whereas CRISPR technology is more specific and sensitive for identifying pathogenic bacteria because it employs programmable CRISPR-Cas systems that target particular DNA sequences, minimizing non-specific binding and cross-reactivity. In this project, a robust detection method based on CRISPR-Cas12a sensing was developed, which is rapid, sensitive and specific for detection of pathogenic E. coli isolates that were collected from the fecal samples from adult goats from 17 farms in Tennessee. Detection reaction contained amplified PCR products for the pathogenic regions, reporter probe, Cas12a enzyme, and crRNA specific to three pathogenic genes-stx1, stx2, and hlyA. The CRISPR reaction with the pathogenic bacteria emitted fluorescence when excited under UV light. To evaluate the detection sensitivity and specificity of this assay, its results were compared with PCR based detection assay. Both methods resulted in similar results for the same samples. This technique is very precise, highly sensitive, quick, cost effective, and easy to use, and can easily overcome the limitations of the present detection methods. This project can result in a versatile detection method that is easily adaptable for rapid response in the detection and surveillance of diseases that pose large-scale biosecurity threats to human health, and plant and animal production.
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  • 文章类型: Journal Article
    背景:SARS-COV-2有越来越多的疾病趋势,因此需要一种快速且负担得起的诊断方法。与其他方法相比,它应该高度准确并节省成本。本研究的目的是实现这些目标。
    方法:本研究使用TaqMan一步RT-qPCR和快速一步RT-LAMP(逆转录酶环介导的等温扩增)分析了342个样品。进行一步LAMP测定以评估灵敏度和特异性。
    结果:研究报告了使用两种不同方法的阳性样本。在RT-LAMP方法中,唾液有92个阳性样本(26.9%)和250个阴性样本(73.09%),鼻咽有94个阳性样本(27.4%)和248个阴性样本(72.51%)。在RT-qPCR方法中,唾液有86个阳性样本(25.1%)和256个阴性样本(74.8%),鼻咽有93个阳性样本(27.1%)和249个阴性样本(72.8%).唾液和鼻咽样本中的两项测试的一致性分别为93%和94%,基于科恩的卡帕系数(κ)(P<0.001)。在1×101的稀释度和100%的特异性下报告了该技术的灵敏度。
    结论:基于研究结果,一步LAMP测定法具有多种优点。这些包括简单性,成本效益,高灵敏度,和特异性。一步LAMP测定法显示出作为诊断工具的希望。它可以帮助管理疾病爆发,确保及时治疗,并通过提供快速,易于使用的测试。
    BACKGROUND: There is an increasing disease trend for SARS-COV-2, so need a quick and affordable diagnostic method. It should be highly accurate and save costs compared to other methods. The purpose of this research is to achieve these goals.
    METHODS: This study analyzed 342 samples using TaqMan One-Step RT-qPCR and fast One-Step RT-LAMP (Reverse Transcriptase Loop-Mediated Isothermal Amplification). The One-Step LAMP assay was conducted to assess the sensitivity and specificity.
    RESULTS: The research reported positive samples using two different methods. In the RT-LAMP method, saliva had 92 positive samples (26.9%) and 250 negative samples (73.09%) and nasopharynx had 94 positive samples (27.4%) and 248 negative samples (72.51%). In the RT-qPCR method, saliva had 86 positive samples (25.1%) and 256 negative samples (74.8%) and nasopharynx had 93 positive samples (27.1%) and 249 negative samples (72.8%). The agreement between the two tests in saliva and nasopharynx samples was 93% and 94% respectively, based on Cohen\'s kappa coefficient (κ) (P < 0.001). The rate of sensitivity in this technique was reported at a dilution of 1 × 101 and 100% specificity.
    CONCLUSIONS: Based on the results of the study the One-Step LAMP assay has multiple advantages. These include simplicity, cost-effectiveness, high sensitivity, and specificity. The One-Step LAMP assay shows promise as a diagnostic tool. It can help manage disease outbreaks, ensure prompt treatment, and safeguard public health by providing rapid, easy-to-use testing.
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  • 文章类型: Journal Article
    多属性方法(MAM)已成为同时筛选治疗性抗体的多种产品质量属性的强大工具。其中一个潜在的关键质量属性(CQA)是糖基化,一种可以影响异质性的常见修改,功能活动,和治疗性抗体的免疫原性。然而,目前监测MAM中糖化水平的方法很少见,而且不够快速和准确.在这项研究中,开发了一种改进的基于质谱(MS)的MAM,以同时监测糖基化和包括非岩藻糖基化在内的其他质量属性.使用具有不同糖基化位点编号的两种治疗性抗体评价该方法。用想法治疗,EndoF2和二硫苏糖醇产生了三个不同的亚基,获得的糖基化结果与用PNGaseF处理的结果相似,这是常规用于释放聚糖;样品处理时间大大减少,同时提供额外的质量属性信息。基于MS的MAM也用于评估在各种缓冲溶液中强制糖化后的糖化进程。当在碳酸氢铵缓冲溶液中进行强制糖化时,观察到氧化的显着增加,共鉴定出23个潜在的糖化位点和4个显著的氧化位点。值得注意的是,我们发现碳酸氢铵能特别刺激氧化,而糖基化对氧化有协同作用。这些发现将这项研究确立为一种新颖的方法,用于实现技术先进的平台和概念,从而增强产品开发和质量控制的功效。其特点是广谱,快速,和准确的性质。
    The multiattribute method (MAM) has emerged as a powerful tool for simultaneously screening multiple product quality attributes of therapeutic antibodies. One such potential critical quality attribute (CQA) is glycation, a common modification that can impact the heterogeneity, functional activity, and immunogenicity of therapeutic antibodies. However, current methods for monitoring glycation levels in MAM are rare and not sufficiently rapid and accurate. In this study, an improved mass spectrometry (MS)-based MAM was developed to simultaneously monitor glycation and other quality attributes including afucosylation. The method was evaluated using two therapeutic antibodies with different glycosylation site numbers. Treatment with IdeS, Endo F2, and dithiothreitol generated three distinct subunits, and the glycation results obtained were similar to those treated with PNGase F, which is routinely used to release glycans; the sample processing time was greatly reduced while providing additional quality attribute information. The MS-based MAM was also employed to assess the glycation progression following forced glycation in various buffer solutions. A significant increase in oxidation was observed when forced glycation was conducted in an ammonium bicarbonate buffer solution, and a total of 23 potential glycation sites and 4 significantly oxidized sites were identified. Notably, we found that ammonium bicarbonate was found to specifically stimulate oxidation, while glycation had a synergistic effect on oxidation. These findings establish this study as a novel methodology for achieving a technologically advanced platform and concept that enhances the efficacy of product development and quality control, characterized by its broad-spectrum, rapid, and accurate nature.
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  • 文章类型: Journal Article
    这么快,使用色谱纸的无设备DNA分离程序是一种简单的方法,可以在不到30分钟的时间内进行,并且不需要湿实验室经验。用最少的费用,它为任何想要探索生物多样性的人提供了一个负担得起的替代方案。它还提供了一个很好的选择,用于教室或其他有时间限制的活动。这种方法最适合植物或地衣,在室温下在Whatman®色谱纸上产生稳定的DNA,可以根据需要洗脱。
    This rapid, equipment-free DNA isolation procedure using chromatography paper is a simple method that can be performed in less than 30 min and requires no wet lab experience. With minimal expense, it offers an affordable alternative for anyone wanting to explore biodiversity. It also provides an excellent option for use in classrooms or other activities that are time limited. The method works best for plants or lichens, producing stable DNA on Whatman® chromatography paper at room temperature, which can be eluted as needed.
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  • 文章类型: Journal Article
    背景:Trio外显子组测序可用于调查妊娠超声发现的先天性异常,但它在澳大利亚的使用尚未被评估。
    目的:评估产前快速基因组检测后的临床结果和管理变化,以指导广泛实施的模型的开发。
    方法:43个全外显子组测序前瞻性转诊,包括40个三人组(父母和怀孕),两名单身人士和一名二人组在三级医院进行了评估,可以使用全州的病理实验室。诊断产量,周转时间(TAT),报告时的胎龄,妊娠结局,对每个家庭的管理变更和未来妊娠状态进行了评估.
    结果:在15/43例妊娠(35%)中进行了具有临床意义的基因组诊断,平均TAT为12天。报告时的妊娠年龄为16+5至31+6周(中位数为21+3周)。分子诊断包括神经肌肉和骨骼疾病,放射病和一系列其他罕见的孟德尔疾病。大多数家庭积极将结果用于怀孕决策以及未来怀孕的管理。
    结论:快速的孕中期产前基因组检测可以成功进行,以调查妊娠的结构异常,为当前和未来的妊娠管理提供重要指导。这种测试的时间敏感性需要密切的实验室和临床合作,以确保适当的转诊和结果沟通。我们发现建立产前协调员角色和专门的报告团队是重要的促进者。我们建议将其作为其他产前服务中基因组测试的模型。
    BACKGROUND: Trio exome sequencing can be used to investigate congenital abnormalities identified on pregnancy ultrasound, but its use in an Australian context has not been assessed.
    OBJECTIVE: Assess clinical outcomes and changes in management after expedited genomic testing in the prenatal period to guide the development of a model for widespread implementation.
    METHODS: Forty-three prospective referrals for whole exome sequencing, including 40 trios (parents and pregnancy), two singletons and one duo were assessed in a tertiary hospital setting with access to a state-wide pathology laboratory. Diagnostic yield, turn-around time (TAT), gestational age at reporting, pregnancy outcome, change in management and future pregnancy status were assessed for each family.
    RESULTS: A clinically significant genomic diagnosis was made in 15/43 pregnancies (35%), with an average TAT of 12 days. Gestational age at time of report ranged from 16 + 5 to 31 + 6 weeks (median 21 + 3 weeks). Molecular diagnoses included neuromuscular and skeletal disorders, RASopathies and a range of other rare Mendelian disorders. The majority of families actively used the results in pregnancy decision making as well as in management of future pregnancies.
    CONCLUSIONS: Rapid second trimester prenatal genomic testing can be successfully delivered to investigate structural abnormalities in pregnancy, providing crucial guidance for current and future pregnancy management. The time-sensitive nature of this testing requires close laboratory and clinical collaboration to ensure appropriate referral and result communication. We found the establishment of a prenatal coordinator role and dedicated reporting team to be important facilitators. We propose this as a model for genomic testing in other prenatal services.
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  • 文章类型: Journal Article
    荷尔蒙皮质醇,作为下丘脑-垂体-肾上腺(HPA)轴的最终产物释放,具有特征良好的昼夜节律,可以对外部压力源产生同种异体反应。当分泌模式被破坏时,皮质醇水平长期升高,导致心脏病等疾病,笔画,精神健康障碍,和糖尿病。慢性应激和应激相关疾病的诊断取决于皮质醇水平的准确测量;目前,它使用质谱或免疫测定进行定量,在有训练有素人员的专业实验室。然而,这些方法很耗时,价格昂贵,无法捕获激素的动态生物节律。这篇重要的评论追溯了皮质醇检测从传统的基于实验室的方法到分散式皮质醇监测生物传感器的路径。提供了皮质醇生物学和病理生理学的完整图片,强调了精准医学方式监测皮质醇的重要性。基于抗体的免疫测定仍然主导着即时生物传感器的开发;新的捕获分子,如适体和分子印迹聚合物(MIP),与微流体等技术相结合,可穿戴电子产品,量子点提供了对检测极限(LoD)的改进,特异性,以及向快速或连续测量的转变。虽然已经提出了各种不同的传感器和设备,目前仍然需要对皮质醇进行定量检测,使用快速或连续的监测设备,这些设备可以实现个性化医疗方法来管理压力。这可以通过可以利用低样本量的技术的协同组合来解决,相关的检测限和快速检测时间,更好地解释皮质醇的生物节律变化。强调了皮质醇诊断朝着激素的快速和连续监测的趋势,以及对样本矩阵选择的见解。
    The hormone cortisol, released as the end-product of the hypothalamic-pituitary-adrenal (HPA) axis, has a well-characterized circadian rhythm that enables an allostatic response to external stressors. When the pattern of secretion is disrupted, cortisol levels are chronically elevated, contributing to diseases such as heart attacks, strokes, mental health disorders, and diabetes. The diagnosis of chronic stress and stress related disorders depends upon accurate measurement of cortisol levels; currently, it is quantified using mass spectroscopy or immunoassay, in specialized laboratories with trained personnel. However, these methods are time-consuming, expensive and are unable to capture the dynamic biorhythm of the hormone. This critical review traces the path of cortisol detection from traditional laboratory-based methods to decentralised cortisol monitoring biosensors. A complete picture of cortisol biology and pathophysiology is provided, and the importance of precision medicine style monitoring of cortisol is highlighted. Antibody-based immunoassays still dominate the pipeline of development of point-of-care biosensors; new capture molecules such as aptamers and molecularly imprinted polymers (MIPs) combined with technologies such as microfluidics, wearable electronics, and quantum dots offer improvements to limit of detection (LoD), specificity, and a shift toward rapid or continuous measurements. While a variety of different sensors and devices have been proposed, there still exists a need to produce quantitative tests for cortisol ─ using either rapid or continuous monitoring devices that can enable a personalized medicine approach to stress management. This can be addressed by synergistic combinations of technologies that can leverage low sample volumes, relevant limit of detection and rapid testing time, to better account for cortisol\'s shifting biorhythm. Trends in cortisol diagnostics toward rapid and continuous monitoring of hormones are highlighted, along with insights into choice of sample matrix.
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