Specialist Antimicrobial Chemotherapy Unit, Public Health Wales Microbiology, Cardiff, CF14 4XW, UK.
Centre for Genomic Pathogen Surveillance, Big Data Institute, University of Oxford, Oxford, OX3 7LF, UK.
Microb Genom. 2023 Sep;9(9). doi: 10.1099/mgen.0.001104.
A multidrug-resistant strain of () sequence type (ST) 1788, an otherwise uncommon ST worldwide, was isolated from 65 patients at 11 hospitals and 11 general practices across South and West Wales, UK, between February 2019 and November 2021. A collection of 97 ST1788 isolates (including 94 from Wales) was analysed to investigate the diversity and spread across Wales and to identify molecular marker(s) to aid development of a strain-specific real-time PCR. Whole genome sequencing (WGS) was performed with Illumina technology and the data were used to perform phylogenetic analyses. Pan-genome analysis of further genome collections was used to identify an ST1788-specific gene target; a real-time PCR was then validated against a panel of 314 strains and 218 broth-enriched screening samples. Low genomic diversity was demonstrated amongst the 94 isolates from Wales. Evidence of spread within and across healthcare facilities was found. A yersiniabactin locus and the KL2 capsular locus were identified in 85/94 (90.4 %) and 94/94 (100 %) genomes respectively; , , and were simultaneously carried by 86/94 (91.5 %) isolates; 4/94 (4.3 %) isolates also carried carbapenemase. Aminoglycoside and fluoroquinolone resistance markers were found in 94/94 (100 %) and 86/94 (91.5 %) isolates respectively. The ST1788-specific real-time PCR was 100 % sensitive and specific. Our analyses demonstrated recent clonal expansion and spread of ST1788 in the community and across healthcare facilities in South and West Wales with isolates carrying well-defined antimicrobial resistance and virulence markers. An ST1788-specific marker was also identified, enabling rapid and reliable preliminary characterization of isolates by real-time PCR. This study confirms the utility of WGS in investigating novel strains and in aiding proactive implementation of molecular tools to assist infection control specialists.
一株多药耐药株的 序列类型 (ST) 1788,一种在全球范围内并不常见的 ST,于 2019 年 2 月至 2021 年 11 月期间,从英国南威尔士和西威尔士的 11 家医院和 11 家综合诊所的 65 名患者中分离出来。对 97 株 ST1788 分离株(包括来自威尔士的 94 株)进行了分析,以调查威尔士的多样性和传播情况,并确定分子标记物以帮助开发针对特定菌株的实时 PCR。采用 Illumina 技术进行全基因组测序 (WGS),并使用数据进行系统发育分析。进一步对 基因组集进行泛基因组分析,以确定 ST1788 特异性基因靶标;然后针对 314 株菌株和 218 个肉汤富集筛选样本对实时 PCR 进行验证。来自威尔士的 94 株分离株的基因组多样性较低。发现了在医疗机构内和之间传播的证据。85/94 (90.4%)和 94/94 (100%)基因组中分别发现了耶尔森菌素基因座和 KL2 荚膜基因座;86/94 (91.5%)分离株同时携带 、 、 和 ;4/94 (4.3%)分离株还携带 碳青霉烯酶。94/94 (100%)和 86/94 (91.5%)分离株均发现氨基糖苷类和氟喹诺酮类耐药标记。ST1788 特异性实时 PCR 的敏感性和特异性均为 100%。我们的分析表明,近期在南威尔士和西威尔士的社区和医疗机构中, ST1788 发生了克隆扩张和传播,分离株携带明确的抗菌和毒力标记物。还确定了 ST1788 特异性标记物,使实时 PCR 能够快速可靠地对分离株进行初步鉴定。本研究证实了 WGS 在调查新型菌株和协助主动实施分子工具以协助感染控制专家方面的实用性。