High throughput

高吞吐量
  • 文章类型: Review
    食物过敏仍然是一种公共卫生,business,和监管挑战。食物过敏原的风险分析(RA)和风险管理(RM)非常重要,对食物过敏原的分析对两者都是必要的。目前用于过敏原分析的主要技术(酶联免疫吸附测定[ELISA]和实时聚合酶链反应)表现出公认的挑战,包括可变和抗体特异性反应以及物种DNA而不是过敏原蛋白的检测。分别。液相色谱-串联质谱(LC-MS/MS)可用于蛋白质鉴定,具有多重分析和可追溯到国际单位制(SI)的潜力,帮助全球测量标准化。在这次审查中,已经系统地回顾了最近的文献,以评估LC-MS/MS的进展,并定义了基质辅助激光解吸/电离飞行时间MS(MALDI-ToF-MS)技术用于过敏原分析的潜力和益处。最初完整蛋白质的MALDI-ToF-MS已经用于验证用于LC-MS/MS分析的计算机衍生的肽序列。我们描述了MALDI的起源及其未来前景,包括与MALDI偶联的亲和珠辅助测定。基于可靠和可重复的基于MALDI的临床应用的增殖,该技术应模仿已建立的过敏原检测技术的检测能力(灵敏度),同时减少技术支持,并具有与竞争技术相当的复用潜力,例如,LC-MS/MS和ELISA。虽然不太可能提供固有的SI可追溯性,基于MALDI的过敏原分析将补充现有的过敏原MS方法。与几乎任何现有技术相比,亲和力珠-MALDI似乎能够以更低的成本实现更高的吞吐量,允许重复子采样,以减少代表性采样问题。
    Food allergy remains a public health, business, and regulatory challenge. Risk analysis (RA) and risk management (RM) of food allergens are of great importance and analysis for food allergens is necessary for both. The current workhorse techniques for allergen analysis (enzyme linked immunosorbent assay [ELISA] and real-time polymerase chain reaction) exhibit recognized challenges including variable and antibody specific responses and detection of species DNA rather than allergen protein, respectively. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) enables protein identification, with potential for multiplex analysis and traceability to the System of International units (SI), aiding global measurement standardization. In this review, recent literature has been systematically reviewed to assess progress in LC-MS/MS and define the potential and benefits of matrix-assisted laser desorption/ionization-time-of-flight MS (MALDI-ToF-MS) technology for allergen analysis. MALDI-ToF-MS of initially intact protein is already applied to verify in silico-derived peptide sequences for LC-MS/MS analysis. We describe the origins of MALDI and its future perspectives, including affinity bead-assisted assays coupled to MALDI. Based on the proliferation of reliable and reproducible MALDI-based clinical applications, the technique should emulate the detection capability (sensitivity) of established allergen detection techniques, whilst reducing technical support and having equivalent multiplexing potential to competing techniques, for example, LC-MS/MS and ELISA. Although unlikely to offer inherent SI traceability, MALDI-based allergen analysis will complement existing MS approaches for allergens. Affinity bead-MALDI appears capable of higher throughput at lower cost per sample than almost any existing technique, enabling repeated sub-sampling as a way to reduce representative sampling issues.
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  • 文章类型: Journal Article
    作为第四大被诊断的癌症,宫颈癌(CC)是影响全球女性的癌症相关死亡率的主要原因之一,特别是在晚期诊断时。CC生物标志物的发现为精准医学铺平了道路,以获得更好的患者预后。高通量组学技术,大数据生产过程进一步加快。迄今为止,通过技术的进步已经发现了各种CC生物标志物。尽管,由于缺乏通过大规模临床研究的验证,很少有人成功转化为临床实践。虽然组学技术产生了大量的数据,在确定转化研究的临床相关数据方面出现了挑战,因为单水平组学方法的分析很少提供因果关系.通过强调所涉及的生物分子及其功能的相互关系,跨不同细胞功能水平的综合多组学方法可以更好地理解CC的基本生物学。帮助识别精准医学的新型综合生物标志物谱。建立全球早期检测研究网络(EDRN)系统有助于加快生物标志物翻译的步伐。为了填补研究空白,从高通量组学技术的应用出发,综述了近年来CC生物标志物研究进展,转录组学,蛋白质组学,和代谢组学。
    As the fourth most diagnosed cancer, cervical cancer (CC) is one of the major causes of cancer-related mortality affecting females globally, particularly when diagnosed at advanced stage. Discoveries of CC biomarkers pave the road to precision medicine for better patient outcomes. High throughput omics technologies, characterized by big data production further accelerate the process. To date, various CC biomarkers have been discovered through the advancement in technologies. Despite, very few have successfully translated into clinical practice due to the paucity of validation through large scale clinical studies. While vast amounts of data are generated by the omics technologies, challenges arise in identifying the clinically relevant data for translational research as analyses of single-level omics approaches rarely provide causal relations. Integrative multi-omics approaches across different levels of cellular function enable better comprehension of the fundamental biology of CC by highlighting the interrelationships of the involved biomolecules and their function, aiding in identification of novel integrated biomarker profile for precision medicine. Establishment of a worldwide Early Detection Research Network (EDRN) system helps accelerating the pace of biomarker translation. To fill the research gap, we review the recent research progress on CC biomarker development from the application of high throughput omics technologies with sections covering genomics, transcriptomics, proteomics, and metabolomics.
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  • 文章类型: Journal Article
    大肠杆菌O157:H7,一种产志贺的大肠杆菌,是一种主要的致病性大肠杆菌菌株,自1980年代初以来已成为重要的食品和水传播病原体。可以应用几种管理策略来控制感染的传播;但是,早期诊断代表了最大程度地减少感染的最佳预防策略。因此,为了降低发病率和死亡率,以快速有效的方式检测这种病原体至关重要。目前使用的金标准测试依赖于来自污染源的大肠杆菌O157:H7的培养和预富集;它们是耗时且费力的。分子方法如聚合酶链反应是敏感的;然而,他们需要昂贵的仪器。因此,有一个准确的要求,敏感,Specific,用户友好,快速,实验室和现场使用的免费设备和可交付(ASSURED)检测方法。等温扩增方法等新兴技术,生物传感器,表面增强拉曼光谱,基于纸的诊断和基于智能手机的数字方法被认为是大肠杆菌O157:H7诊断领域的新方法,并在这篇综述中进行了讨论。移动PCR和CRISPR-Cas诊断平台已被确定为大肠杆菌O157:H7POC诊断的新工具,具有在行业中实施的潜力。本文综述了在食品和水工业背景下大肠杆菌O157:H7诊断领域的进展和进展。重点是新兴的高通量护理点(POC)大肠杆菌O157:H7诊断以及食品和水行业常规诊断服务转型的要求。
    Escherichia coli O157:H7, a Shiga-producing E. coli is a major pathogenic E. coli strain which since the early 1980s has become a crucial food and water-borne pathogen. Several management strategies can be applied to control the spread of infection; however early diagnosis represents the optimum preventive strategy to minimize the infection. Therefore, it is crucial to detect this pathogen in a fast and efficient manner in order to reduce the morbidity and mortality. Currently used gold standard tests rely on culture and pre-enrichment of E. coli O157:H7 from the contaminated source; they are time consuming and laborious. Molecular methods such as polymerase chain reaction are sensitive; however, they require expensive instrumentation. Therefore, there is a requirement for Accurate, Sensitive, Specific, User friendly, Rapid, Equipment free and Deliverable (ASSURED) detection methods for use in the laboratory and in the field. Emerging technologies such as isothermal amplification methods, biosensors, surface enhanced Raman Spectroscopy, paper-based diagnostics and smartphone-based digital methods are recognized as new approaches in the field of E. coli O157:H7 diagnostics and are discussed in this review. Mobile PCR and CRISPR-Cas diagnostic platforms have been identified as new tools in E. coli O157:H7 POC diagnostics with the potential for implementation by industry. This review describes advances and progress in the field of E. coli O157:H7 diagnosis in the context of food and water industry. The focus is on emerging high throughput point-of-care (POC) E. coli O157:H7 diagnostics and the requirement for the transformation to service routine diagnostics in the food and water industry.
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  • 文章类型: Journal Article
    The review aimed to identify the different high-throughput phenotyping (HTP) techniques that used for quality evaluation in cassava and yam breeding programmes, and this has provided insights towards the development of metrics and their application in cassava and yam improvements. A systematic review of the published research articles involved the use of NIRS in analysing the quality traits of cassava and yam was carried out, and Scopus, Science Direct, Web of Sciences and Google Scholar were searched. The results of the review established that NIRS could be used in understanding the chemical constituents (carbohydrate, protein, vitamins, minerals, carotenoids, moisture, starch, etc.) for high-throughput phenotyping. This study provides preliminary evidence of the application of NIRS as an efficient and affordable procedure for HTP. However, the feasibility of using mid-infrared spectroscopy (MIRS) and hyperspectral imaging (HSI) in combination with the NIRS could be further studied for quality traits phenotyping.
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  • 文章类型: Journal Article
    As a malignant tumor type, nasopharyngeal carcinoma (NPC) is characterized by distinct geographical, ethnic and genetic differences; presenting a major threat to human health in many countries, especially in Southern China. At present, no accurate and effective methods are available for the early diagnosis, efficacious evaluation or prognosis prediction for NPC. As such, a large number of patients have locoregionally advanced NPC at the time of initial diagnosis. Many patients show toxic reactions to overtreatment and have risks of cancer recurrence and distant metastasis owing to insufficient treatment. To solve these clinical problems, high‑throughput \'‑omics\' technologies are being used to screen and identify specific molecular biomarkers for NPC. Because of the lack of comprehensive descriptions regarding NPC biomarkers, the present study summarized the research progress that has been made in recent years to discover NPC biomarkers, highlighting the existing problems that require exploration. In view of the lack of authoritative reports at present, study design factors that affect the screening of biomarkers are also discussed here and prospects for future research are proposed to provide references for follow‑up studies of NPC biomarkers.
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  • 文章类型: Journal Article
    Microplastic research is a rapidly developing field, with urgent needs for high throughput and automated analysis techniques. We conducted a review covering image analysis from optical microscopy, scanning electron microscopy, fluorescence microscopy, and spectral analysis from Fourier transform infrared (FT-IR) spectroscopy, Raman spectroscopy, pyrolysis gas-chromatography mass-spectrometry, and energy dispersive X-ray spectroscopy. These techniques were commonly used to collect, process, and interpret data from microplastic samples. This review outlined and critiques current approaches for analysis steps in image processing (color, thresholding, particle quantification), spectral processing (background and baseline subtraction, smoothing and noise reduction, data transformation), image classification (reference libraries, morphology, color, and fluorescence intensity), and spectral classification (reference libraries, matching procedures, and best practices for developing in-house reference tools). We highlighted opportunities to advance microplastic data analysis and interpretation by (i) quantifying colors, shapes, sizes, and surface topologies with image analysis software, (ii) identifying threshold values of particle characteristics in images that distinguish plastic particles from other particles, (iii) advancing spectral processing and classification routines, (iv) creating and sharing robust spectral libraries, (v) conducting double blind and negative controls, (vi) sharing raw data and analysis code, and (vii) leveraging readily available data to develop machine learning classification models. We identified analytical needs that we could fill and developed supplementary information for a reference library of plastic images and spectra, a tutorial for basic image analysis, and a code to download images from peer reviewed literature. Our major findings were that research on microplastics was progressing toward the use of multiple analytical methods and increasingly incorporating chemical classification. We suggest that new and repurposed methods need to be developed for high throughput screening using a diversity of approaches and highlight machine learning as one potential avenue toward this capability.
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  • 文章类型: Journal Article
    The significance of thermodynamic solubility in biopharmaceutical compound or drug characterization as well as the importance of having methods that accurately establish it have been extensively addressed. Nonetheless, its precise determination continues to remain a challenging task to accomplish. Even more so when the number of compounds to evaluate is high and the available amount of each compound is low, both of which are inevitable for the compound characterization during the drug development process. Except for the shake-flask method which is still considered as the \'gold standard\' in obtaining thermodynamic data, it is currently difficult to say that another satisfactory model which is routinely used to determine thermodynamic solubility is being applied. Therefore, this review summarizes the various experimental approaches which are based on the classical shake flask method but have yet attempted to speed up the experimental process of obtaining such data more conveniently. The most important experimental features of these approaches are provided to the reader. Some advantages and disadvantages associated with each approach are also highlighted, consequently offering a resource to those looking for the most appropriate of the approaches that have already fared well at determining the biopharmaceutically relevant drug solubility.
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  • 文章类型: Journal Article
    Data processing and analysis are major bottlenecks in high-throughput metabolomic experiments. Recent advancements in data acquisition platforms are driving trends toward increasing data size (e.g., petabyte scale) and complexity (multiple omic platforms). Improvements in data analysis software and in silico methods are similarly required to effectively utilize these advancements and link the acquired data with biological interpretations. Herein, we provide an overview of recently developed and freely available metabolomic tools, algorithms, databases, and data analysis frameworks. This overview of popular tools for MS and NMR-based metabolomics is organized into the following sections: data processing, annotation, analysis, and visualization. The following overview of newly developed tools helps to better inform researchers to support the emergence of metabolomics as an integral tool for the study of biochemistry, systems biology, environmental analysis, health, and personalized medicine.
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  • 文章类型: Journal Article
    优化使用木质纤维素生物质作为可再生能源生产的原料目前正在全球范围内开发。生物质是纤维素的复杂混合物,半纤维素,木质素,提取物,和蛋白质;以及无机盐。用于生物质表征的细胞壁组成分析是费力且耗时的。为了快速有效地表征生物量,已经成功开发了几种高通量技术。其中,近红外光谱(NIR)和热解分子束质谱(Py-mbms)是互补的工具,能够在短时间内评估大量的原始或改性生物质。NIR显示与特定化学结构相关的振动,而Py-mbms描绘了来自生物质分解的全部片段。NIR振动和Py-mbms峰都属于可能的化学官能团和分子结构。它们提供了生物材料化学见解的补充信息。然而,由于光谱中包含大量的重叠带或分解片段,因此解释信息结果具有挑战性。为了提高数据分析的效率,多变量分析工具已经被用来定义数据变量之间的显著相关性,使得大量的带/峰可以被表示原始变化的少量重构变量代替。重建的数据变量用于样品比较(主成分分析)和在生物质化学结构和感兴趣的性质之间建立回归模型(偏最小二乘回归)。在这次审查中,总结了NIR和Py-mbms测得的重要生物质化学结构。介绍了常规数据分析方法和多元数据分析方法的优缺点,比较和评价。这篇综述旨在为选择最有效的NIR和Py-mbms表征生物量的数据分析方法提供指导。
    Optimizing the use of lignocellulosic biomass as the feedstock for renewable energy production is currently being developed globally. Biomass is a complex mixture of cellulose, hemicelluloses, lignins, extractives, and proteins; as well as inorganic salts. Cell wall compositional analysis for biomass characterization is laborious and time consuming. In order to characterize biomass fast and efficiently, several high through-put technologies have been successfully developed. Among them, near infrared spectroscopy (NIR) and pyrolysis-molecular beam mass spectrometry (Py-mbms) are complementary tools and capable of evaluating a large number of raw or modified biomass in a short period of time. NIR shows vibrations associated with specific chemical structures whereas Py-mbms depicts the full range of fragments from the decomposition of biomass. Both NIR vibrations and Py-mbms peaks are assigned to possible chemical functional groups and molecular structures. They provide complementary information of chemical insight of biomaterials. However, it is challenging to interpret the informative results because of the large amount of overlapping bands or decomposition fragments contained in the spectra. In order to improve the efficiency of data analysis, multivariate analysis tools have been adapted to define the significant correlations among data variables, so that the large number of bands/peaks could be replaced by a small number of reconstructed variables representing original variation. Reconstructed data variables are used for sample comparison (principal component analysis) and for building regression models (partial least square regression) between biomass chemical structures and properties of interests. In this review, the important biomass chemical structures measured by NIR and Py-mbms are summarized. The advantages and disadvantages of conventional data analysis methods and multivariate data analysis methods are introduced, compared and evaluated. This review aims to serve as a guide for choosing the most effective data analysis methods for NIR and Py-mbms characterization of biomass.
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  • 文章类型: Journal Article
    Gas chromatography is widely applied to separate, identify, and quantify components of samples in a timely manner. Increasing demand for analytical throughput, instrument portability, environmental sustainability, and more economical analysis necessitates the development of new gas chromatography instrumentation. The applications of resistive column heating technologies have been espoused for nearly thirty years and resistively heated gas chromatography has been commercially available for the last ten years. Despite this lengthy period of existence, resistively heated gas chromatography has not been universally adopted. This low rate of adoption may be partially ascribed to the saturation of the market with older convection oven technology, coupled with other analytical challenges such as sampling, injection, detection and data processing occupying research. This article assesses the advantages and applications of resistive heating in gas chromatography and discusses practical considerations associated with adoption of this technology.
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