Schuster H J, van Mansfeld R, van der Reijden W A, van Houdt R, Matamoros S
Medical Microbiology and Infection Control, Amsterdam UMC Location University of Amsterdam, Amsterdam, the Netherlands; Amsterdam Institute for Infection and Immunity, Amsterdam UMC Location University of Amsterdam, Amsterdam, the Netherlands.
Laboratory of Medical Microbiology, OLVG Lab, Amsterdam, the Netherlands.
J Hosp Infect. 2025 Aug;162:351-359. doi: 10.1016/j.jhin.2025.04.021. Epub 2025 May 6.
Outbreaks with vancomycin-resistant Enterococcus faecium (VRE) are common in hospitals worldwide. Whole-genome MLST (wgMLST) is often used to identify outbreak strains, but VRE typing can still be challenging due to their limited genomic variation.
Developing a method for sequence analysis of vancomycin resistance genes in parallel with wgMLST and application of this new method for real-time investigation of two parallel VRE outbreaks.
A bioinformatics pipeline was developed to compare the sequences of transposons containing vanB resistance genes. This pipeline was used in addition to wgMLST to investigate two separate ongoing VRE outbreaks. Five separate colonies from 15 different samples and 10 vancomycin-susceptible isolates were also sequenced.
Of 46 strains collected during two outbreaks, 26 and nine strains were identified as being part of the two outbreaks based on wgMLST clustering. In six strains an identical vanB transposon but a different wgMLST cluster were identified, indicating horizontal gene transfer. This potential outbreak spread would have been missed without transposon analysis. There was no evidence of variability in vanB transposon sequence or wgMLST profiles within different colonies from the same sample. One vancomycin-susceptible E. faecium in blood culture was identified, with a wgST similar to one of the outbreak strains.
Real-time analysis of transposons containing vancomycin resistance genes provides additional information for analysis of vanB-VRE outbreaks. It detects possible horizontal gene transfer which would not be detected using conventional methods. Transposon analysis is a valuable addition to whole-genome sequence analysis during vanB-VRE outbreaks.
耐万古霉素屎肠球菌(VRE)暴发在全球医院中很常见。全基因组多位点序列分型(wgMLST)常用于鉴定暴发菌株,但由于VRE的基因组变异有限,其分型仍具有挑战性。
开发一种与wgMLST并行的万古霉素耐药基因序列分析方法,并将这种新方法应用于实时调查两起并行的VRE暴发。
开发了一种生物信息学流程,用于比较含有vanB耐药基因的转座子序列。除wgMLST外,该流程还用于调查两起独立的正在发生的VRE暴发。还对来自15个不同样本的5个独立菌落和10株万古霉素敏感分离株进行了测序。
在两次暴发期间收集的46株菌株中,基于wgMLST聚类,分别有26株和9株被鉴定为两次暴发的一部分。在6株菌株中,鉴定出相同的vanB转座子但wgMLST聚类不同,表明存在水平基因转移。如果没有转座子分析,这种潜在的暴发传播可能会被遗漏。在来自同一样本的不同菌落中,没有证据表明vanB转座子序列或wgMLST图谱存在变异性。血培养中鉴定出一株万古霉素敏感屎肠球菌,其wgST与其中一株暴发菌株相似。
对含有万古霉素耐药基因的转座子进行实时分析为vanB-VRE暴发分析提供了额外信息。它能检测到使用传统方法无法检测到的可能的水平基因转移。在vanB-VRE暴发期间,转座子分析是全基因组序列分析的一项有价值的补充。