| RETRACTED: Paper Lateral Flow Biosensor for Nodavirus Reverse Transcribed RNA Detection
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Author:
Date:
2020-08-05
[Abstract] Paper nanobiosensors have been established as an excellent platform for analysis of veterinary and human pathogens causing various diseases. Especially, lateral flow assays or biosensors ideal for sensitive, rapid, robust and accurate analysis in laboratory setups and on-site analysis. Viral RNA detection is of great importance for public health as well as animal health protection. In that aspect, the present protocol focuses on the development of functionalized gold nanoparticle-based lateral flow biosensor for fish nervous necrosis virus (Nodavirus) nucleic acids detection. Total viral RNA, isolated from fish samples was subjected to reverse transcription PCR amplification and the amplification products were mixed with specific oligonucleotide probe. A red test line was formed when ...
[摘要] [摘要] 纸纳米生物传感器已经成为分析导致各种疾病的兽医和人类病原体的一个极好的平台。尤其是侧向流分析或生物传感器是实验室设备和现场分析中灵敏、快速、可靠和准确分析的理想选择。病毒RNA检测对公共卫生和动物健康保护具有重要意义。在这一方面，本协议的重点是开发功能化金纳米粒子侧流生物传感器，用于鱼类神经坏死病毒（Nodavirus）核酸检测。从鱼类标本中分离出的总病毒RNA进行逆转录PCR扩增，扩增产物与特异性寡核苷酸探针混合。当有nodavirus产物存在时，形成红色检测线。这种方法对基础研究有很大的意义，因为它消除了耗时、繁琐的电泳程序的需要，并且可以在养鱼场对养鱼户进行调整。利用这种生物分析平台进行病害监测，无需耗费大量时间和成本，对水产养殖和环境安全有很大影响。
[背景] 关注点和/或现场生物分析一直是关注人类和动物福祉的研究工作的最终目标。基于纸基的传感平台具有功能化简单、重现性好、制造成本低等优点，是一种极具吸引力的分析平台。纸基分析设备已应用于小分子、蛋白质和各种核酸的分析（Parolo和Merkoçi，2013；Bahadir和Sezgintürk，2016；Jiang等人，2019）。侧流生物传感器（LFB）是一种带有干试剂的预制材料条带，通过流体样品激活。它们专为一次性一次性使用而设计，只要有足够的开/关信号（Posthuma ...
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| RNA Extraction from Ears and Draining Lymph Nodes of Mice Infected with Leishmania amazonensis
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Author:
Date:
2020-06-05
[Abstract] Parasites of the genus Leishmania infect the mammalian hosts, including mice and humans and cause cutaneous or visceral leishmaniasis depending upon the parasite species transmitted by the vector sandfly. Leishmania amazonensis is one of the Leishmania species responsible for the cutaneous form of the disease. We have inoculated with these parasites the ear dermis of mice. RNA preparations were performed from fragmented tissues using a buffer containing guanidin isothiocynate (RLT buffer, RNeasy Mini Kit, Qiagen, SAS, France) and β-mercaptoethanol. Both reagents facilitate the isolation of intact RNA from tissues and the use of the RNeasy Kits present with several advantages that facilitate the isolation of pure non-degraded total RNA: i) This method allows to ...
[摘要] [摘要] 寄生虫 利什曼原虫感染哺乳动物宿主，包括小鼠和人类的原因皮肤或内脏Leishm Aniasis根据在转交了该载体的寄生虫种类白蛉。利什曼原虫亚马孙是的利什曼原虫物种负责对皮肤型疾病。我们已经接种这些RNA制备物从零散组织用含有缓冲液中进行guanidin 异硫氰酸（RLT缓冲液，RNeasy Mini试剂盒，Qiagen公司，SAS，法国）和β - 巯基乙醇。 两种试剂促进完整的RNA的从组织中分离，并与促进纯非恶化的总RNA的分离几个优点使用RNeasy试剂试剂盒本的：我）Ť 他的方法允许避免苯酚的在RNA提取存在缓冲液，通常在备选方案中使用;ⅱ）此外焦碳酸二乙酯（DEPC）处理的玻璃器皿，以避免RNA酶的样品的污染，不与该协议所需的;ⅲ）˚F inally，它是一个快速的过程和分离的总RNA可将其浓缩在小体积中，以利于其用于下游实验程序。
[背景] 属的病原体利什曼出生动物传染病与多种临床表现和疾病严重程度可变性的原因矢量（曼索托等人，2007）利什曼病的皮肤.The形式是由几个引起利什曼原虫种包括利什曼原虫亚马孙（麦克马洪-Pratt and Alexander，2004; Christensen et al。，2019）。
小鼠模型的存在，对皮肤利什曼病的敏感性高，中度或无敏感性，促进了寄生虫传播实验规程的发展（de ...
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| Rapid Generation of Human Neuronal Cell Models Enabling Inducible Expression of Proteins-of-interest for Functional Studies
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Author:
Date:
2020-05-05
[Abstract] CRISPR-Cas9 technology has transformed the ability to edit genomic sequences and control gene expression with unprecedented ease and scale. However, precise genomic insertions of coding sequences using this technology remain time-consuming and inefficient because they require introducing adjacent single-strand cuts through Cas9 nickase action and invoking the host-encoded homology-directed repair program through the concomitant introduction of large repair templates. Here, we present a system for the rapid study of any protein-of-interest in two neuronal cell models following its inducible expression from the human AAVS1 safe harbor locus. With lox-flanked foundation cassettes in the AAVS1 site and a tailor-made plasmid for accepting coding sequences-of-interest in ...
[摘要] [摘要] CRISPR-Cas9技术以前所未有的简便性和规模改变了编辑基因组序列和控制基因表达的能力。但是，由于需要引入相邻的单链，因此使用该技术进行精确的基因组编码插入仍然很耗时且效率低下。通过减少Cas9切口酶的作用并通过同时引入大型修复模板来调用宿主编码的同源性指导的修复程序。在此，我们提出了一种系统，用于在其诱导后的两个神经元细胞模型中快速研究任何目的蛋白该系统可从人类AAVS1 安全港基因座表达，在AAVS1 位点具有lox侧翼的基础盒和定制的质粒以接受感兴趣的编码序列，该系统使研究人员能够为诱导型产生自己的神经元细胞模型任何编码表达序列不到一个月的时间。由于可用性Preinserted 增强型绿色的Fluo 可以与目标蛋白质融合的最新蛋白质（EGFP）编码序列，该系统可帮助功能研究通过活细胞显微镜以及利用非常有效的可用性进行的相互作用组分析来跟踪目标蛋白质EGFP捕获矩阵。
[背景] ...
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