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比较下一代技术和生物信息学管道在荚膜分型中的应用。

Comparison of next generation technologies and bioinformatics pipelines for capsular typing of .

机构信息

Department of RDI Microbiology, Hospital Sant Joan de Déu , Barcelona, Spain.

Infectious Diseases and Microbiome, Institut de Recerca Sant Joan de Déu , Barcelona, Spain.

出版信息

J Clin Microbiol. 2023 Dec 19;61(12):e0074123. doi: 10.1128/jcm.00741-23. Epub 2023 Nov 21.

Abstract

Whole genome sequencing (WGS)-based approaches for pneumococcal capsular typing have become an alternative to serological methods. serotyping from WGS has not yet been applied to long-read sequences produced by third-generation technologies. The objective of the study was to determine the capsular types of pneumococci causing invasive disease in Catalonia (Spain) using serological typing and WGS and to compare the performance of different bioinformatics pipelines using short- and long-read data from WGS. All invasive pneumococcal pediatric isolates collected in Hospital Sant Joan de Déu (Barcelona) from 2013 to 2019 were included. Isolates were assigned a capsular type by serological testing based on anticapsular antisera and by different WGS-based pipelines: Illumina sequencing followed by serotyping with PneumoCaT, SeroBA, and Pathogenwatch vs MinION-ONT sequencing coupled with serotyping by Pathogenwatch from pneumococcal assembled genomes. A total of 119 out of 121 pneumococcal isolates were available for sequencing. Twenty-nine different serotypes were identified by serological typing, with 24F ( = 17; 14.3%), 14 ( = 10; 8.4%), and 15B/C ( = 8; 6.7%) being the most common serotypes. WGS-based pipelines showed initial concordance with serological typing (>91% of accuracy). The main discrepant results were found at the serotype level within a serogroup: 6A/B, 6C/D, 9A/V, 11A/D, and 18B/C. Only one discrepancy at the serogroup level was observed: serotype 29 by serological testing and serotype 35B/D by all WGS-based pipelines. Thus, bioinformatics WGS-based pipelines, including those using third-generation sequencing, are useful for pneumococcal capsular assignment. Possible discrepancies between serological typing and WGS-based approaches should be considered in pneumococcal capsular-type surveillance studies.

摘要

基于全基因组测序(WGS)的方法已成为血清学方法的替代方法,用于肺炎球菌荚膜分型。然而,WGS 血清分型尚未应用于第三代技术产生的长读序列。本研究的目的是使用血清学分型和 WGS 确定加泰罗尼亚(西班牙)侵袭性疾病的肺炎球菌荚膜型,并比较使用 WGS 的短读和长读数据的不同生物信息学管道的性能。纳入了 2013 年至 2019 年在巴塞罗那圣若翰医院采集的所有侵袭性小儿肺炎球菌分离株。通过血清学检测基于抗荚膜抗血清和不同的 WGS 为基础的管道,将分离株分配一个荚膜型:Illumina 测序后用 PneumoCaT、SeroBA 和 Pathogenwatch 进行血清型鉴定,以及 MinION-ONT 测序后用 Pathogenwatch 从肺炎球菌组装基因组进行血清型鉴定。共有 119 株肺炎球菌分离株可用于测序。通过血清学分型鉴定出 29 种不同的血清型,其中 24F(=17;14.3%)、14(=10;8.4%)和 15B/C(=8;6.7%)最为常见。基于 WGS 的管道与血清学分型具有初始一致性(>91%的准确性)。在血清型水平上,在一个血清群内发现了主要的不一致结果:6A/B、6C/D、9A/V、11A/D 和 18B/C。仅在血清群水平上观察到一个不一致结果:血清型 29 经血清学检测,所有基于 WGS 的管道均为血清型 35B/D。因此,包括第三代测序在内的基于生物信息学 WGS 的管道可用于肺炎球菌荚膜归属。在肺炎球菌荚膜型监测研究中,应考虑血清学分型和基于 WGS 的方法之间的可能差异。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3310/10729682/8e2febc8f302/jcm.00741-23.f001.jpg

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