Bioinformatics Unit (MF 1), Department for Methods Development and Research Infrastructure, Robert Koch Institute, 13353 Berlin, Germany.
Centre for Biological Threats and Special Pathogens, Highly Pathogenic Viruses (ZBS1), Robert Koch Institute, 13353 Berlin, Germany.
Viruses. 2019 Jun 8;11(6):536. doi: 10.3390/v11060536.
Emerging virus diseases present a global threat to public health. To detect viral pathogens in time-critical scenarios, accurate and fast diagnostic assays are required. Such assays can now be established using mass spectrometry-based targeted proteomics, by which viral proteins can be rapidly detected from complex samples down to the strain-level with high sensitivity and reproducibility. Developing such targeted assays involves tedious steps of peptide candidate selection, peptide synthesis, and assay optimization. Peptide selection requires extensive preprocessing by comparing candidate peptides against a large search space of background proteins. Here we present Purple (Picking unique relevant peptides for viral experiments), a software tool for selecting target-specific peptide candidates directly from given proteome sequence data. It comes with an intuitive graphical user interface, various parameter options and a threshold-based filtering strategy for homologous sequences. Purple enables peptide candidate selection across various taxonomic levels and filtering against backgrounds of varying complexity. Its functionality is demonstrated using data from different virus species and strains. Our software enables to build taxon-specific targeted assays and paves the way to time-efficient and robust viral diagnostics using targeted proteomics.
新兴病毒疾病对全球公共卫生构成威胁。为了在时间紧迫的情况下及时检测病毒病原体,需要准确、快速的诊断检测方法。现在,基于质谱的靶向蛋白质组学可以建立这样的检测方法,通过该方法可以从复杂的样本中以高灵敏度和重现性快速检测到病毒蛋白,甚至可以精确到株水平。开发这种靶向检测方法涉及到繁琐的步骤,包括肽候选物的选择、肽合成和检测优化。肽选择需要通过将候选肽与大量背景蛋白的搜索空间进行广泛的预处理来完成。在这里,我们介绍了 Purple(用于病毒实验的特异性肽选择),这是一种从给定蛋白质组序列数据中直接选择目标特异性肽候选物的软件工具。它具有直观的图形用户界面,各种参数选项和基于阈值的同源序列过滤策略。Purple 可以跨各种分类水平进行肽候选物选择,并可以针对不同复杂程度的背景进行过滤。我们使用来自不同病毒物种和株的数据演示了其功能。我们的软件可以构建分类特异性靶向检测方法,为使用靶向蛋白质组学进行高效、稳健的病毒诊断铺平了道路。