{"title":"临床微生物实验室细菌分型的全自动全基因组测序平台。","authors":"Rachel L Siddall, Jordan C Starkey, Robin Patel","doi":"10.1128/jcm.00178-25","DOIUrl":null,"url":null,"abstract":"<p><p>Nosocomial outbreaks impact patient safety and place an economic burden on healthcare facilities. Laboratory testing plays a crucial role in outbreak investigation and guides containment efforts. In recent years, whole genome sequencing (WGS) methods have replaced traditional typing methods due to higher resolution and simplified workflows. Though an improvement from previous methods, bacterial WGS is time-consuming with manual DNA extraction and library preparation and long sequencing times for paired-end sequence data. Here, a fully automated library preparation and sequencing solution, the Clear Dx<sup>TM</sup> WGS platform (Clear Labs, San Carlos, CA), was compared to a more manual library preparation and sequencing approach using 226 isolates representing 18 bacterial species. Sequence data were analyzed using SeqSphere+ (Ridom, Münster, Germany), and the results of the two methods were compared. Of the 224 isolate sequences analyzed, 222 (99%) showed concordant isolate groupings, and, overall, the results of the two approaches were statistically similar by comparison of distance matrices. The automated workflow reduced turnaround time by 16-19 h and eliminated 3 h of manual labor while decreasing costs by an estimated 34%-57% depending on the number of isolates run. This study demonstrates the advantages of integrating automation into bacterial WGS workflows.IMPORTANCEAn automated platform for bacterial nucleic acid extraction and whole genome sequencing was compared to a manual method for bacterial strain typing. The two approaches yielded nearly equivalent results, with the automated approach providing improvement in turnaround time and cost, with less manual pipetting.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":"63 5","pages":"e0017825"},"PeriodicalIF":6.1000,"publicationDate":"2025-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12077132/pdf/","citationCount":"0","resultStr":"{\"title\":\"Automated whole genome sequencing platform for bacterial strain typing in clinical microbiology laboratories.\",\"authors\":\"Rachel L Siddall, Jordan C Starkey, Robin Patel\",\"doi\":\"10.1128/jcm.00178-25\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Nosocomial outbreaks impact patient safety and place an economic burden on healthcare facilities. Laboratory testing plays a crucial role in outbreak investigation and guides containment efforts. 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The automated workflow reduced turnaround time by 16-19 h and eliminated 3 h of manual labor while decreasing costs by an estimated 34%-57% depending on the number of isolates run. This study demonstrates the advantages of integrating automation into bacterial WGS workflows.IMPORTANCEAn automated platform for bacterial nucleic acid extraction and whole genome sequencing was compared to a manual method for bacterial strain typing. 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引用次数: 0
摘要
院内疫情影响患者安全,并给医疗机构带来经济负担。实验室检测在疫情调查和指导控制工作中起着至关重要的作用。近年来,全基因组测序(WGS)方法以更高的分辨率和简化的工作流程取代了传统的分型方法。虽然与以前的方法相比有所改进,但细菌WGS需要人工提取DNA和制备文库,耗时长,对端序列数据的测序时间长。在这里,我们将全自动文库制备和测序解决方案Clear DxTM WGS平台(Clear Labs, San Carlos, CA)与更手动的文库制备和测序方法进行了比较,使用了226株代表18种细菌的菌株。序列数据采用SeqSphere+ (Ridom, m nster, Germany)进行分析,并比较两种方法的结果。224个分离物序列中,222个(99%)分离物分类一致,总体上,两种方法的结果在距离矩阵比较上具有统计学上的相似性。自动化工作流程减少了16-19小时的周转时间,消除了3小时的人工劳动,同时根据运行的分离次数,成本降低了34%-57%。这项研究证明了将自动化集成到细菌WGS工作流程中的优势。对细菌核酸提取和全基因组测序的自动化平台与菌株分型的人工方法进行了比较。这两种方法产生了几乎相同的结果,自动化方法提供了周转时间和成本的改进,减少了人工移液。
Automated whole genome sequencing platform for bacterial strain typing in clinical microbiology laboratories.
Nosocomial outbreaks impact patient safety and place an economic burden on healthcare facilities. Laboratory testing plays a crucial role in outbreak investigation and guides containment efforts. In recent years, whole genome sequencing (WGS) methods have replaced traditional typing methods due to higher resolution and simplified workflows. Though an improvement from previous methods, bacterial WGS is time-consuming with manual DNA extraction and library preparation and long sequencing times for paired-end sequence data. Here, a fully automated library preparation and sequencing solution, the Clear DxTM WGS platform (Clear Labs, San Carlos, CA), was compared to a more manual library preparation and sequencing approach using 226 isolates representing 18 bacterial species. Sequence data were analyzed using SeqSphere+ (Ridom, Münster, Germany), and the results of the two methods were compared. Of the 224 isolate sequences analyzed, 222 (99%) showed concordant isolate groupings, and, overall, the results of the two approaches were statistically similar by comparison of distance matrices. The automated workflow reduced turnaround time by 16-19 h and eliminated 3 h of manual labor while decreasing costs by an estimated 34%-57% depending on the number of isolates run. This study demonstrates the advantages of integrating automation into bacterial WGS workflows.IMPORTANCEAn automated platform for bacterial nucleic acid extraction and whole genome sequencing was compared to a manual method for bacterial strain typing. The two approaches yielded nearly equivalent results, with the automated approach providing improvement in turnaround time and cost, with less manual pipetting.
期刊介绍:
The Journal of Clinical Microbiology® disseminates the latest research concerning the laboratory diagnosis of human and animal infections, along with the laboratory's role in epidemiology and the management of infectious diseases.