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SYBR 绿 I/碘化丙啶法快速检测临床分离大肠埃希菌的抗生素敏感性。

Rapid determination of antibiotic susceptibility of clinical isolates of Escherichia coli by SYBR green I/Propidium iodide assay.

机构信息

Department of Clinical Laboratory of Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, Shandong, China.

Institute of Clinical Microbiology, Shandong Academy of Clinical Medicine, Jinan, Shandong, China.

出版信息

Sci Rep. 2024 Aug 13;14(1):18782. doi: 10.1038/s41598-024-69286-7.

Abstract

Infections caused by pathogenic Escherichia coli are a serious threat to human health, while conventional antibiotic susceptibility tests (AST) have a long turn-around time, and rapid antibiotic susceptibility methods are urgently needed to save lives in the clinic, reduce antibiotic misuse and prevent emergence of antibiotic-resistant bacteria. We optimized and validated the feasibility of a novel rapid AST based on SYBR Green I and Propidium Iodide (SGPI-AST) for E. coli drug susceptibility test. A total of 112 clinical isolates of E. coli were collected and four antibiotics (ceftriaxone, cefoxitin, imipenem, meropenem) were selected for testing. Bacterial survival rate of E. coli was remarkably linearly correlated with S value at different OD600 values. After optimizing the antibiotic concentrations, the sensitivity and specificity of SGPI-AST reached 100%/100%, 97.8%/100%, 100%/100% and 98.4%/99% for ceftriaxone, cefoxitin, imipenem and meropenem, respectively, and the corresponding concordances of the SGPI-AST with conventional AST were 1.000, 0.980, 1.000 and 0.979, respectively. The SGPI-AST can rapidly and accurately determine the susceptibility of E. coli clinical isolates to multiple antibiotics in 60 min, and has the potential to be applied to guide the precise selection of antibiotics for clinical management of infections caused by pathogenic E. coli.

摘要

产毒性大肠杆菌引起的感染对人类健康构成严重威胁,而传统的抗生素药敏试验(AST)耗时较长,因此临床上急需快速的抗生素药敏方法,以拯救生命、减少抗生素滥用和预防抗生素耐药菌的出现。我们优化并验证了基于 SYBR Green I 和碘化丙啶(SGPI-AST)的新型快速 AST 用于大肠杆菌药敏试验的可行性。共收集了 112 株临床分离的大肠杆菌,选择了 4 种抗生素(头孢曲松、头孢西丁、亚胺培南、美罗培南)进行检测。不同 OD600 值下大肠杆菌的存活率与 S 值呈显著线性相关。优化抗生素浓度后,SGPI-AST 对头孢曲松、头孢西丁、亚胺培南和美罗培南的灵敏度和特异性分别达到 100%/100%、97.8%/100%、100%/100%和 98.4%/99%,与传统 AST 的一致性分别为 1.000、0.980、1.000 和 0.979。SGPI-AST 可在 60 分钟内快速准确地确定大肠杆菌临床分离株对多种抗生素的药敏性,有望应用于指导临床管理产毒性大肠杆菌感染时抗生素的精准选择。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8bb7/11322298/6f0dcacbacbf/41598_2024_69286_Fig1_HTML.jpg

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