Pathology and LaboratoryMedicine, David Geffen School of Medicine, University of California, Los Angeles.
Mi Next Generation Science Core Laboratory, Children's Hospital Los Angeles, California.
Clin Infect Dis. 2017 Apr 1;64(7):894-901. doi: 10.1093/cid/ciw876.
Whole-genome sequencing (WGS) is an emerging and powerful technique by which to perform epidemiological studies in outbreak situations.
WGS was used to identify and evaluate an outbreak of OXA-232-expressing carbapenem-resistant Klebsiella pneumoniae (CRKP) transmitted to 16 patients over the course of 40 weeks via endoscopic retrograde cholangiopancreatography procedures at a single institution. WGS was performed on 32 OXA-232 CRKP isolates (1-7 per patient) and single-nucleotide variants (SNVs) were analyzed, with reference to the index patient's isolate.
Interhost genetic diversity of isolates was between 0 and 15 SNVs during the outbreak; molecular clock calculations estimated 12.31 substitutions per genome per year (95% credibility interval, 7.81-17.05). Both intra- and interpatient diversification at the plasmid and transposon level was observed, significantly impacting the antibiogram of outbreak isolates. The majority of isolates evaluated (n = 27) harbored a blaCTX-M-15 gene, but some (n = 5) lacked the transposon carrying this gene, which resulted in susceptibility to aztreonam and third- and fourth-generation cephalosporins. Similarly, an isolate from a colonized patient lacked the transposon carrying rmtF and aac(6')lb genes, resulting in susceptibility to aminoglycosides.
This study broadens the understanding of how bacteria diversify at the genomic level over the course of a defined outbreak and provides reference for future outbreak investigations.
全基因组测序(WGS)是一种新兴的强大技术,可用于在暴发情况下进行流行病学研究。
WGS 用于鉴定和评估在一家机构进行的 40 周内镜逆行胰胆管造影术过程中,通过内镜逆行胰胆管造影术传播给 16 名患者的 OXA-232 表达碳青霉烯类耐药肺炎克雷伯菌(CRKP)暴发。对 32 株 OXA-232 CRKP 分离株(每位患者 1-7 株)进行 WGS,并分析单核苷酸变异(SNV),并参考索引患者的分离株。
暴发期间,分离株之间的宿主间遗传多样性为 0 至 15 SNV;分子钟计算估计每年每个基因组有 12.31 个替换(95%置信区间,7.81-17.05)。在质粒和转座子水平上观察到了种内和种间的多样化,这显著影响了暴发分离株的药敏谱。评估的大多数分离株(n=27)携带 blaCTX-M-15 基因,但有些分离株(n=5)缺乏携带该基因的转座子,这导致对氨曲南和第三代和第四代头孢菌素的敏感性。同样,定植患者的分离株缺乏携带 rmtF 和 aac(6')lb 基因的转座子,导致对氨基糖苷类药物的敏感性。
本研究拓宽了对细菌在特定暴发过程中基因组水平多样化的理解,并为未来的暴发调查提供了参考。