Development of a PCR assay for rapid and accurate detection of an emerging vanB Enterococcus faecium clone in the Capital Region of Denmark.

IF 3.7 Q2 INFECTIOUS DISEASES JAC-Antimicrobial Resistance Pub Date : 2024-11-07 eCollection Date: 2024-12-01 DOI:10.1093/jacamr/dlae180

Maja Johanne Søndergaard Knudsen, Christel Barker Jensen, Rikke Lind Jørgensen, Andreas Munk Petersen, Gitte Qvist Kristiansen, Jan Gorm Lisby, Peder Worning, Henrik Westh, Mette Pinholt

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Abstract

Objectives: To develop and validate a real-time PCR assay detecting the sequence bridging Tn1549 and the Enterococcus faecium chromosome in the emerging vanB vancomycin-resistant E. faecium (VREfm) clone (ST80/CT2406).

Methods: The Tn1549 insertion site was determined on routinely sequenced VREfm isolates. The outer boundaries of Tn1549 and adjoining host bacterial sequences were determined using a BLAST search in the silent information regulator gene sir2. Next, the primers and probe were developed, targeting the sequence bridging Tn1549 and the E. faecium chromosome. Finally, the PCR assay was validated on well-characterized strains and prospectively performed on rectal screening samples submitted to our laboratory.

Results and conclusions: The PCR assay proved to be accurate and provide rapid diagnosis of the emerging vanB VREfm in rectal screening samples.

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开发用于快速准确检测丹麦首都地区新出现的 vanB 粪肠球菌克隆的 PCR 检测方法。

目的开发并验证一种实时 PCR 检测方法，以检测新出现的耐万古霉素粪肠球菌（VanB）克隆（ST80/CT2406）中 Tn1549 与粪肠球菌染色体的桥接序列：方法：在常规测序的 VREfm 分离物中确定了 Tn1549 插入位点。然后，针对连接 Tn1549 和粪肠球菌染色体的序列开发了引物和探针。最后，PCR 检测在特征明确的菌株上进行了验证，并对提交给我们实验室的直肠筛查样本进行了前瞻性检测：结果和结论：PCR 检测法被证明是准确的，能快速诊断直肠筛查样本中新出现的 vanB VREfm。

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