PMA

PMa
  • 文章类型: Journal Article
    衣壳完整性定量PCR(qPCR),一种将偶氮染料预处理与qPCR相结合的感染性病毒分子检测方法,近年来已被广泛使用;然而,各种病毒类型的预处理条件的变化可能会限制特定方案的效力。通过在过去十年(2009-2019年)中在食品安全和环境病毒学领域对病毒进行衣壳完整性qPCR的鉴定和严格综合了最近的41项同行评审研究,我们旨在建立检测传染性病毒的建议。嵌入染料是有效的措施,在PCR测定生存力的病毒衣壳被破坏;已经被其他原因灭活的病毒,例如丧失依恋或基因组损伤,使用这种方法不太容易被检测到。尽管建议为每种病毒优化特定的协议,我们确定了一般测定条件的框架。这些包括单叠氮乙锭的浓度,单叠氮丙啶或其衍生物在10至200μM之间;在冰上或在室温(20-25°C)下孵育5-120分钟;并使用LED或强光(500-800瓦)暴露5至20分钟的时间进行染料激活。这些简单的步骤可以有助于在常规(水)监测环境中以及在病毒暴发期间(例如当前的COVID-19大流行或登革热等地方病)进行传染性病毒传播的调查。
    Capsid integrity quantitative PCR (qPCR), a molecular detection method for infectious viruses combining azo dye pretreatment with qPCR, has been widely used in recent years; however, variations in pretreatment conditions for various virus types can limit the efficacy of specific protocols. By identifying and critically synthesizing forty-one recent peer-reviewed studies employing capsid integrity qPCR for viruses in the last decade (2009-2019) in the fields of food safety and environmental virology, we aimed to establish recommendations for the detection of infectious viruses. Intercalating dyes are effective measures of viability in PCR assays provided the viral capsid is damaged; viruses that have been inactivated by other causes, such as loss of attachment or genomic damage, are less well detected using this approach. Although optimizing specific protocols for each virus is recommended, we identify a framework for general assay conditions. These include concentrations of ethidium monoazide, propidium monoazide or its derivates between 10 and 200 μM; incubation on ice or at room temperature (20 - 25 °C) for 5-120 min; and dye activation using LED or high light (500-800 Watts) exposure for periods ranging from 5 to 20 min. These simple steps can benefit the investigation of infectious virus transmission in routine (water) monitoring settings and during viral outbreaks such as the current COVID-19 pandemic or endemic diseases like dengue fever.
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  • 文章类型: Journal Article
    Mass spectrometry (MS) is the most versatile and comprehensive method in \"OMICS\" sciences (i.e. in proteomics, genomics, metabolomics and lipidomics). The applications of MS and tandem MS (MS/MS or MS(n)) provide sequence information of the full complement of biological samples in order to understand the importance of the sequences on their precise and specific functions. Nowadays, the control of polymer sequences and their accurate characterization is one of the significant challenges of current polymer science. Therefore, a similar approach can be very beneficial for characterizing and understanding the complex structures of synthetic macromolecules. MS-based strategies allow a relatively precise examination of polymeric structures (e.g. their molar mass distributions, monomer units, side chain substituents, end-group functionalities, and copolymer compositions). Moreover, tandem MS offer accurate structural information from intricate macromolecular structures; however, it produces vast amount of data to interpret. In \"OMICS\" sciences, the software application to interpret the obtained data has developed satisfyingly (e.g. in proteomics), because it is not possible to handle the amount of data acquired via (tandem) MS studies on the biological samples manually. It can be expected that special software tools will improve the interpretation of (tandem) MS output from the investigations of synthetic polymers as well. Eventually, the MS/MS field will also open up for polymer scientists who are not MS-specialists. In this review, we dissect the overall framework of the MS and MS/MS analysis of synthetic polymers into its key components. We discuss the fundamentals of polymer analyses as well as recent advances in the areas of tandem mass spectrometry, software developments, and the overall future perspectives on the way to polymer sequencing, one of the last Holy Grail in polymer science.
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