Bratcher Holly B, Harrison Odile B, Maiden Martin C J
Department of Zoology, University of Oxford, Oxford, UK.
Methods Mol Biol. 2019;1969:51-82. doi: 10.1007/978-1-4939-9202-7_4.
Whole genome sequencing (WGS) has revolutionized molecular microbiology, allowing the population biology of bacterial pathogens to be examined with greater accuracy and detail. The study of Neisseria meningitidis isolates, in particular, has benefitted from the availability of WGS data allowing outbreak cases, hyper-invasive lineages, molecular epidemiology, and vaccine coverage to be determined. Here, we describe a suite of protocols for the optimum recovery and analysis of WGS data, including a brief overview of methods for N. meningitidis DNA extraction, sequencing, and analysis. Downstream analysis tools are described including a step-by-step guide to the use of PubMLST.org/neisseria . This freely accessible website provides a resource for the Neisseria community allowing the diversity of the meningococcal population to be extracted and exploited.
全基因组测序(WGS)彻底改变了分子微生物学,使人们能够更准确、更详细地研究细菌病原体的群体生物学。特别是,脑膜炎奈瑟菌分离株的研究受益于WGS数据的可用性,这些数据可用于确定暴发病例、高侵袭性谱系、分子流行病学和疫苗覆盖率。在此,我们描述了一套用于最佳恢复和分析WGS数据的方案,包括对脑膜炎奈瑟菌DNA提取、测序和分析方法的简要概述。还介绍了下游分析工具,包括使用PubMLST.org/neisseria的分步指南。这个免费访问的网站为奈瑟菌研究群体提供了一个资源,使人们能够提取和利用脑膜炎球菌群体的多样性。