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采用富集接种肉汤直接检测 vanB 型万古霉素耐药肠球菌的 Xpert vanA/vanB 检测法评估。

Evaluation of the Xpert vanA/vanB assay using enriched inoculated broths for direct detection of vanB vancomycin-resistant Enterococci.

机构信息

Department of Medical Microbiology, University of Groningen, University Medical Center Groningen, Groningen, the Netherlands

Department of Medical Microbiology, University of Groningen, University Medical Center Groningen, Groningen, the Netherlands.

出版信息

J Clin Microbiol. 2014 Dec;52(12):4293-7. doi: 10.1128/JCM.01125-14. Epub 2014 Oct 8.

Abstract

Rapid and accurate detection of VRE (vancomycin-resistant enterococci) is required for adequate antimicrobial treatment and infection prevention measures. Previous studies using PCR for the detection of VRE, including Cepheid's Xpert vanA/vanB assay, reported accurate detection of vanA VRE; however, many false-positive results were found for vanB VRE. This is mainly due to nonenterococcal vanB genes, which can be found in the gut flora. Our goal was to optimize the rapid and accurate detection of vanB VRE and to improve the positive predictive value (PPV) by limiting false-positive results. We evaluated the use of the Xpert vanA/vanB assay on rectal swabs and on enriched inoculated broths for the detection of vanB VRE. By adjusting the cycle threshold (CT) cutoff value to ≤ 25 for positivity by PCR on enriched broths, the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were 96.9%, 100%, 100%, and 99.5% for vanB VRE, respectively. As shown in this study, CT values of ≤ 25 acquired from enriched broths can be considered true positive. For broths with CT values between 25 and 30, we recommend confirming the results by culture. CT values of >30 appeared to be true negative. In conclusion, this study shows that the Cepheid's Xpert vanA/vanB assay performed on enriched inoculated broths with an adjusted cutoff CT value is a useful and rapid tool for the detection of vanB VRE.

摘要

快速准确地检测万古霉素耐药肠球菌(VRE)对于适当的抗菌治疗和感染预防措施至关重要。之前使用 PCR 检测 VRE 的研究,包括 Cepheid 的 Xpert vanA/vanB 检测,报告了 vanA VRE 的准确检测;然而,vanB VRE 存在许多假阳性结果。这主要是由于非肠球菌的 vanB 基因可存在于肠道菌群中。我们的目标是优化 vanB VRE 的快速准确检测,并通过限制假阳性结果来提高阳性预测值(PPV)。我们评估了 Xpert vanA/vanB 检测在直肠拭子和富集接种肉汤中检测 vanB VRE 的用途。通过将 CT 值(PCR 在富集肉汤中检测阳性的截止值)调整为≤25,vanB VRE 的灵敏度、特异性、阳性预测值(PPV)和阴性预测值(NPV)分别为 96.9%、100%、100%和 99.5%。如本研究所示,来自富集肉汤的 CT 值≤25 可被认为是真正的阳性。对于 CT 值在 25 和 30 之间的肉汤,我们建议通过培养确认结果。CT 值>30 似乎为真正的阴性。总之,本研究表明,Cepheid 的 Xpert vanA/vanB 检测在调整后的 CT 值富集接种肉汤上进行是一种有用且快速的检测 vanB VRE 的工具。

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