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产碳青霉烯酶的耐碳青霉烯菌的快速筛查:使用肉汤增菌直肠拭子进行免疫层析试验的临床应用

Rapid screening for carbapenemase-producing carbapenem-resistant : clinical implementation of an immunochromatographic test using broth-enriched rectal swabs.

作者信息

P A M Bezerra Camila Loredana, Castro Barbara de Almeida Lessa, Levin Anna Sara, Baldi Lindissy Luara, Cury Ana Paula, Rossi Flavia, Reese Natashia, Lessa Fernanda C, Bollinger Susan, Salomão Matias Chiarastelli

机构信息

Department of Infectious Diseases, Faculdade de Medicina, Universidade de São Paulo, São Paulo, Brazil.

Department of Infectious Diseases, Faculdade de Medicina, Hospital das Clínicas, Universidade de São Paulo, São Paulo, Brazil.

出版信息

Microbiol Spectr. 2025 Sep 2;13(9):e0131325. doi: 10.1128/spectrum.01313-25. Epub 2025 Aug 8.

Abstract

Carbapenemase-producing carbapenem-resistant (CP-CRE) poses a public health issue. Rapid detection of CP-CRE colonization is challenging; existing methods are either expensive or time-consuming. We evaluated an immunochromatographic test (ICT) for detecting carbapenemases directly from broth-enriched rectal swabs. One hundred intensive care patients provided 178 pairs of rectal swabs. One swab was tested using the GeneXpert Carba-R PCR assay; the other was inoculated into brain-heart infusion broth. After 4 and 6 h of incubation at 37°C, the broth was tested with the RESIST-5 O.K.N.V.I. ICT for carbapenemase (KPC), New Delhi metallo-β-lactamase (NDM), oxacillinase-48 (OXA-48), Verona integron-encoded metallo-β-lactamase (VIM), and imipenemase (IMP) carbapenemases. Broths were subcultured after overnight incubation, and recovered carbapenem-resistant isolates were tested using GeneXpert Carba-R PCR. Sensitivity, specificity, and accuracy were calculated in comparison to culture positive for CP-CRE, confirmed by PCR for KPC, NDM, OXA-48, VIM, and IMP. Of the 178 swabs, 60 were culture positive for CP-CRE. After 6 h, the ICT demonstrated a sensitivity of 65%, specificity of 97.5%, and accuracy of 86.8%. Among heavily soiled swabs, sensitivity reached 81.8% for ICT after 6 h, and the specificity was 100%. The mean execution time for carbapenemase detection using ICT was reduced by 60 h compared to culture. The ICT after 6 h incubation offers reduced execution time for detecting CP-CREs. This method may serve as a valuable rapid screening tool, especially in resource-limited settings.IMPORTANCEThe rapid spread of multidrug-resistant bacteria requires innovative solutions for early detection and prevention measures. In this study, we present a simple protocol for the direct detection of carbapenemases in rectal swabs using an immunochromatographic assay. By optimizing the assay conditions, we achieved rapid and high-accuracy identification of five clinically important carbapenemases. This method can broaden access to rapid CP-CRE detection of fecal colonization-even in laboratories with limited resources-enabling the implementation of faster and more effective infection control measures, potentially reducing the spread of resistance.

摘要

产碳青霉烯酶的耐碳青霉烯肠杆菌科细菌(CP-CRE)构成了一个公共卫生问题。快速检测CP-CRE定植具有挑战性;现有方法要么昂贵,要么耗时。我们评估了一种免疫层析试验(ICT),用于直接从肉汤增菌的直肠拭子中检测碳青霉烯酶。100名重症监护患者提供了178对直肠拭子。一对拭子中的一个使用GeneXpert Carba-R PCR检测法进行检测;另一个接种到脑心浸液肉汤中。在37°C孵育4小时和6小时后,用RESIST-5 O.K.N.V.I. ICT检测肉汤中的碳青霉烯酶(KPC)、新德里金属β-内酰胺酶(NDM)、氧青霉烷酶-48(OXA-48)、维罗纳整合子编码金属β-内酰胺酶(VIM)和美罗培南酶(IMP)。过夜孵育后将肉汤进行传代培养,对分离出的耐碳青霉烯菌使用GeneXpert Carba-R PCR进行检测。与经KPC、NDM、OXA-48、VIM和IMP的PCR确认的CP-CRE培养阳性结果相比,计算敏感性、特异性和准确性。在178份拭子中,60份CP-CRE培养呈阳性。6小时后,ICT的敏感性为65%,特异性为97.5%,准确性为86.8%。在污染严重的拭子中,6小时后ICT的敏感性达到81.8%,特异性为100%。与培养法相比,使用ICT检测碳青霉烯酶的平均执行时间缩短了60小时。孵育6小时后的ICT检测CP-CRE的执行时间缩短。该方法可作为一种有价值的快速筛查工具,尤其是在资源有限的环境中。重要性多重耐药菌的迅速传播需要创新的早期检测和预防措施解决方案。在本研究中,我们提出了一种使用免疫层析试验直接检测直肠拭子中碳青霉烯酶的简单方案。通过优化试验条件,我们实现了对五种临床上重要的碳青霉烯酶的快速、高精度鉴定。这种方法可以扩大粪便定植中快速CP-CRE检测的可及性,即使在资源有限的实验室中也是如此,从而能够实施更快、更有效的感染控制措施,有可能减少耐药性的传播。

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