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Crystal Violet


Company: Sigma-Aldrich
Catalog#: C0775
Other protocol()

Colorimetric RT-LAMP Methods to Detect Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2)
[Abstract]  Standard diagnostic methods of Coronavirus Disease 2019 (COVID-19) rely on RT-qPCR technique which have limited point-of-care test (POCT) potential due to necessity of dedicated equipment and specialized personnel. LAMP, an isothermal nucleic acid amplification test (NAAT), is a promising technique that may substitute RT-qPCR for POCT of genomic materials. Here, we provide a protocol to perform reverse transcription LAMP targeting SARS-CoV-2. We adopted both real-time fluorescence detection and end-point colorimetric detection approaches. Our protocol would be useful for screening diagnosis of COVID-19 and be a baseline for development of improved POCT NAAT. [摘要]  [摘要] 2019年冠状病毒病的标准诊断方法(COVID-19)依靠RT-qPCR技术,由于需要专用设备和专门人员,因此具有有限的即时检验(POCT)潜力。LAMP是一种等温核酸扩增测试(NAAT),是一种有前途的技术,可以用RT-qPCR代替基因组材料的POCT。在这里,我们提供了一种执行针对SARS-CoV-2的逆转录LAMP的协议。我们同时采用了实时荧光检测和终点比色检测方法。我们的协议对于筛查COVID-19的诊断将是有用的,并且是开发改进的POCT NAAT的基线。

[背景]快速和SARS-CoV的-2,COVID-19的病原的灵敏检测,重要的是要控制电流大流行的情况,因为它使得早期检测,隔离和治疗以及监测筛选。按照世界卫生组织的建议,当前用于检测SARS-CoV-2的标准方法采用RT-qPCR (世界卫生组织,2020年)。然而,RT-qPCR诊断的正确执行需要高水平的设施和专家,通常在医院/采样场所无法获得,因此缺乏POCT适用性。

由于等温扩增方法可以用相对简单的仪器进行检测,因此开发了等温扩增方法以通过在各种现场即时检测核酸来完成诊断(Niemz et ...

In vitro Detection of Neutrophil Traps and Post-attack Cell Wall Changes in Candida Hyphae
[Abstract]  In this protocol we describe how to visualize neutrophil extracellular traps (NETs) and fungal cell wall changes in the context of the coculture of mouse neutrophils with fungal hyphae of Candida albicans. These protocols are easily adjusted to test a wide array of hypotheses related to the impact of immune cells on fungi and the cell wall, making them promising tools for exploring host-pathogen interactions during fungal infection. [摘要]  在本协议中,我们描述了如何在小鼠嗜中性粒细胞与真菌白色念珠菌的真菌菌丝共培养的背景下观察嗜中性粒细胞胞外捕获物(NETs)和真菌细胞壁变化。 这些方案很容易调整,以测试与免疫细胞对真菌和细胞壁的影响有关的各种假设,使其成为探索真菌感染期间宿主病原体相互作用的有前景的工具。

C。白色念珠菌是一种多态性机会性酵母,嗜中性粒细胞是免疫细胞,对于这种和其他真菌病原体的防御至关重要(Brown等,2012; Lionakis和Netea,2013)。 NETs是可以针对病原体部署的潜在防御机制,并且已经表明它们优先针对诸如C的微生物细胞部署。白念珠菌菌丝太大而不能吞噬(Urban et al。,2006; Bruns等人,2010; Branzk等人,,2014; Rohm等人,2014)。 NET已经显示含有许多组分,包括髓过氧化物酶,胞外DNA和瓜氨酸化的组蛋白(Amulic等人,2012; ...

Human Blood Component Vaccinia Virus Neutralization Assay
[Abstract]  Many therapeutic viruses, such as oncolytic viruses, vaccines, or gene therapy vectors, may be administered by the intravenous route to maximize their delivery to target tissues. Blood components, such as antibody, complement and blood cells (such as neutrophils, monocytes, T cells, B cells or platelets) may result in viral neutralization and therefore reduce the therapeutic efficacy. This protocol will describe an in vitro assay by which to test the interaction of viruses with blood components. The effect of various factors can be isolated through fractionation. While whole blood can offer the most physiologically relevant snapshot, plasma can investigate the effects of antibody in concert with complement, and heat inactivated plasma will interrogate the effect of antibody ... [摘要]  许多治疗性病毒,例如溶瘤病毒,疫苗或基因治疗载体,可以通过静脉内途径施用以最大化它们向靶组织的递送。 血液组分,例如抗体,补体和血细胞(例如嗜中性粒细胞,单核细胞,T细胞,B细胞或血小板)可导致病毒中和并因此降低治疗功效。 该方案将描述用于测试病毒与血液组分的相互作用的体外测定。 各种因素的影响可以通过分馏分离。 尽管全血可以提供最具生理学相关性的快照,但血浆可以研究抗体与补体一起的作用,而热灭活的血浆将会询问单独的抗体的作用。