Screening for vancomycin-resistant enterococci: an efficient and economical laboratory-developed test
Screening for vancomycin-resistant enterococci: an efficient and economical laboratory-developed test
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DOI：
10.1007/s10096-011-1304-0
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发表时间：
2012-03-01
影响因子：
4.5
通讯作者：
Ullberg, M.
中科院分区：
文献类型：
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作者：
Fang, H.;Ohlsson, A. -K.;Ullberg, M.
本研究开发并评估了一种实验室开发的测试（实验室测定），结合了增菌肉汤和实时聚合酶链反应 (PCR)，用于筛选万古霉素耐药肠球菌 (VRE)。通过 Lab Assay 和基于 Roche LightCycler VRE 检测试剂盒的方法，对总共 1,765 份粪便或直肠拭子送往实验室进行 VRE 筛查进行并行研究。实验室检测的诊断值如下：100% 灵敏度、79.92% 特异性、1.94% 阳性预测值和 100% 阴性预测值，与基于 LightCycler 试剂盒的检测结果相当。对于 VanA 型和 VanB 型 VRE，实验室测定的检测限均为 10(0) 至 10(1) 菌落形成单位 (CFU)/ml 肉汤接种物。本研究开发的 PCR 方法被批准适用于 Applied Biosystems 7500 快速实时 PCR 系统和 LightCycler(A(R)) 480 实时 PCR 系统。选择 PCR 系统的灵活性使得 PCR 检测可以与 DNA 提取平台完全兼容，从而提供集成的工作流程。此外，在我们的 VRE 常规筛查中，当实验室检测取代基于商业试剂盒的方法时，每个样品的材料成本可节省 7 欧元。因此，实验室开发的肉汤 PCR 方法是一种有效且经济的 VRE 筛查方法。
A laboratory-developed test (Lab Assay), combining enrichment broth and real-time polymerase chain reaction (PCR) for vancomycin-resistant enterococci (VRE) screening, was developed and evaluated in this study. A total of 1,765 faecal or rectal swabs sent to the laboratory for VRE screening were investigated in parallel by Lab Assay and the Roche LightCycler VRE detection kit-based method. The diagnostic values for Lab Assay were as follows: 100% sensitivity, 79.92% specificity, 1.94% positive predictive value and 100% negative predictive value, which were comparable to the results from the LightCycler kit-based assay. The detection limit of Lab Assay was 10(0) to 10(1) colony-forming units (CFU)/ml of inoculum in broth for both VanA-type and VanB-type VRE. The PCR method developed in this study was approved to be applicable on both the Applied Biosystems 7500 Fast Real-Time PCR System and the LightCycler(A (R)) 480 Real-Time PCR System. The flexibility in choosing PCR systems makes it possible that the PCR assay could be fully compatible with the DNA extraction's platform, providing an integrated workflow. Furthermore, the material cost is saved at 7EUR per sample when Lab Assay replaces the commercial kit-based method in our routine screening for VRE. Therefore, the laboratory-developed broth-PCR method is an efficient and economical assay for VRE screening.