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基于实验室监测百日咳博德特氏菌的短读长全基因组测序

Short-Read Whole-Genome Sequencing for Laboratory-Based Surveillance of Bordetella pertussis.

作者信息

Marchand-Austin Alex, Tsang Raymond S W, Guthrie Jennifer L, Ma Jennifer H, Lim Gillian H, Crowcroft Natasha S, Deeks Shelley L, Farrell David J, Jamieson Frances B

机构信息

Department of Laboratory Medicine and Pathobiology, Faculty of Medicine, University of Toronto, Toronto, Canada

Public Health Ontario Laboratory, Public Health Ontario, Toronto, Canada.

出版信息

J Clin Microbiol. 2017 May;55(5):1446-1453. doi: 10.1128/JCM.02436-16. Epub 2017 Feb 22.

Abstract

is a Gram-negative bacterium that causes respiratory infections in humans. Ongoing molecular surveillance of acellular vaccine (aP) antigens is critical for understanding the interaction between evolutionary pressures, disease pathogenesis, and vaccine effectiveness. Methods currently used to characterize aP components are relatively labor-intensive and low throughput. To address this challenge, we sought to derive aP antigen genotypes from minimally processed short-read whole-genome sequencing data generated from 40 clinical isolates and analyzed using the SRST2 bioinformatic package. SRST2 was able to identify aP antigen genotypes for all antigens with the exception of pertactin, possibly due to low read coverage in GC-rich low-complexity regions of variation. Two main genotypes were observed in addition to a singular third genotype that contained an 84-bp deletion that was identified by SRST2 despite the issues in allele calling. This method has the potential to generate large pools of molecular data that can be linked to clinical and epidemiological information to facilitate research of vaccine effectiveness and disease severity in the context of emerging vaccine antigen-deficient strains.

摘要

是一种可导致人类呼吸道感染的革兰氏阴性细菌。对无细胞疫苗(aP)抗原进行持续的分子监测对于理解进化压力、疾病发病机制和疫苗效力之间的相互作用至关重要。目前用于表征aP成分的方法相对劳动强度大且通量低。为应对这一挑战,我们试图从40株临床分离株产生的经过最少处理的短读长全基因组测序数据中推导aP抗原基因型,并使用SRST2生物信息学软件包进行分析。SRST2能够识别除百日咳杆菌黏附素之外的所有抗原的aP抗原基因型,这可能是由于富含GC的低复杂度变异区域的读长覆盖度较低。除了一种单一的第三种基因型外,还观察到两种主要基因型,该第三种基因型包含一个84碱基对的缺失,尽管在等位基因分型方面存在问题,但SRST2仍识别出了该缺失。这种方法有可能生成大量的分子数据,这些数据可与临床和流行病学信息相关联,以促进在新出现的疫苗抗原缺陷菌株背景下对疫苗效力和疾病严重程度的研究。

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