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使用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)和Fc-MBL@FeO富集技术检测玻璃体液中金黄色葡萄球菌的耐甲氧西林情况。

Detection of methicillin resistance of Staphylococcus aureus in vitreous humor using MALDI-TOF MS and Fc-MBL@FeO enrichment.

作者信息

Ren Jun, Gao Wenjing, Yu Menghuan, Liu Chunhong, Ding Chuanfan, Li Shengjie, Yu Shaoning, Cao Wenjun

机构信息

Clinical Laboratory, Eye & ENT Hospital, Fudan University, Shanghai, 200031, China.

Institute of Mass Spectrometry, School of Material Science and Chemical Engineering, Ningbo University, Ningbo, 315211, Zhejiang, China.

出版信息

Sci Rep. 2024 Dec 28;14(1):30757. doi: 10.1038/s41598-024-80715-5.

Abstract

Endophthalmitis is a serious infectious eye disease that causes permanent vision loss. This study developed a method for rapid identification and drug resistance analysis of pathogens in vitreous humor. After short-term rapid culture, 30 Staphylococcus aureus isolates were enriched and purified from the vitreous humor using Fc-MBL@FeO, and then identified by MALDI-TOF MS. The bacterial solution was adjusted to 10 CFU/mL and mixed with CAMHB containing cefoxitin (4 µg/mL) at the same volume. After culture, it was enriched by Fc-MBL@FeO and identified by MALDI-TOF MS. MRSA was judged according to whether the bacteria could successfully be identified. The enrichment efficiency of Fc-MBL@FeO for S. aureus in CAMHB was 88.1%. The detection rate of S. aureus reached 100% after 8 h of vitreous humor culture. The best test performance was achieved with Fc-MBL@FeO enrichment after a 3 h incubation. At this time point, 96.7% validity, 100% sensitivity, and 100% specificity were achieved. Thus, the identification and drug resistance analysis of S. aureus (51-110 CFU) in vitreous humor was completed within 11 h. This study provides a new method for rapid clinical diagnosis of endophthalmitis and precise treatment with antibiotics.

摘要

眼内炎是一种严重的感染性眼病,可导致永久性视力丧失。本研究开发了一种用于快速鉴定玻璃体液中病原体并进行耐药性分析的方法。经过短期快速培养,使用Fc-MBL@FeO从玻璃体液中富集并纯化出30株金黄色葡萄球菌分离株,然后通过基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)进行鉴定。将菌液调整至10 CFU/mL,并与含头孢西丁(4 μg/mL)的CAMHB以相同体积混合。培养后,用Fc-MBL@FeO进行富集,并通过MALDI-TOF MS进行鉴定。根据细菌能否成功鉴定来判断耐甲氧西林金黄色葡萄球菌(MRSA)。Fc-MBL@FeO对CAMHB中金黄色葡萄球菌的富集效率为88.1%。玻璃体液培养8小时后金黄色葡萄球菌的检出率达到100%。孵育3小时后用Fc-MBL@FeO富集可获得最佳检测性能。此时,有效性为96.7%,灵敏度和特异性均为100%。因此,在11小时内完成了玻璃体液中金黄色葡萄球菌(51-110 CFU)的鉴定和耐药性分析。本研究为眼内炎的快速临床诊断和抗生素精准治疗提供了一种新方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eec0/11681162/0d5ff8ad38bb/41598_2024_80715_Fig1_HTML.jpg

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