淋病是我国乃至世界最常见的性传播疾病，其病原体淋球菌的耐药性作为严重的公共卫生问题受到世界卫生组织的重点关注。为了描述淋球菌的耐药及分布特征，需要同时测序其20余个耐药与分型基因，因此，如使用现有的Sanger测序方法，则要大量的重复实验和手工数据分析，并且必须以分离到菌株为前提，费时费力。为解决上述问题，本研究中我们设想应用MinION纳米孔测序仪，结合我们前期工作中已掌握的MinION实验及数据分析流程，使用多重PCR扩增子测序的策略，建立一种准确、灵敏、特异、快速的新型淋球菌耐药及分型基因测序方法。新的方法可自临床样本中同时扩增20余个目标基因并完成测序，通过设计生物信息学分析流程，自动获得目的基因的完整序列。单张芯片可以分析24个临床样本。相对于现有方法，新方法不需要进行菌株的分离培养，极大减少了实验及数据分析的工作量。此外，该方法的建立流程还可作为标准化的范式外推到其他重要病原体

Gonorrhea remains the most significant sexually transmitted disease in China as well as the world. Neisseria gonorrhoeae (gonococcus), as the etiological agent of gonorrhea, is kept under constant surveillance by WHO because its progressively serious resistance. To comprehensively describe its characteristics of antimicrobial resistance (AMR) and distribution, more than 20 genes related to AMR and typing should be sequenced. Sanger sequencing is the most commonly used method, however, substantial repeated experiments and manual data analyzing make it a laborious task. Moreover, prior preparation of bacterial isolation is indispensable, which is also time-consuming. Therefore, this study aims to develop a novel method for rapidly sequencing genes related to AMR and typing in N. gonorrhoeae with high accuracy, sensitivity and specificity. The novel method can simultaneously amplify above 20 targeting genes directly from clinical samples and subsequently obtain complete sequences using the bioinformatics workflow designed specifically for this method. About 24 different samples can be analyzed within one flowcell. Compared with Sanger sequencing, without bacterial culture and repeatedly manual work, the new method can reduce the workload dramatically. Additionally, the established process has the potential of being extended to other important pathogens, thus providing new strategies for studies of molecular epidemiology of infectious diseases.