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[评估选择性显色琼脂培养基(chromID VRE)和多重聚合酶链反应方法检测耐万古霉素肠球菌的实用性]

[Evaluation of the usefulness of selective chromogenic agar medium (chromID VRE) and multiplex PCR method for the detection of vancomycin-resistant enterococci].

作者信息

Kim Do-Hoon, Lee Jae-Hee, Ha Jung-Sook, Ryoo Nam-Hee, Jeon Dong-Seok, Kim Jae-Ryong

机构信息

Department of Laboratory Medicine, Keimyung University School of Medicine, Daegu, Korea.

出版信息

Korean J Lab Med. 2010 Dec;30(6):631-6. doi: 10.3343/kjlm.2010.30.6.631.

Abstract

BACKGROUND

Accurate and early detection of vancomycin-resistant enterococci (VRE) is critical for controlling nosocomial infection. In this study, we evaluated the usefulness of a selective chromogenic agar medium and of multiplex PCR for detection of VRE, and both these techniques were compared with the conventional culture method for VRE detection.

METHODS

We performed the following 3 methods for detecting VRE infection in stool specimens: the routine culture method, culturing in selective chromogenic agar medium (chromID VRE, bioMérieux, France), and multiplex PCR using the Seeplex® VRE ACE Detection kit (Seegene Inc., Korea) with additional PCR for vanC genes.

RESULTS

We isolated 109 VRE strains from 100 stool specimens by the routine culture method. In chromID VRE, all the isolates showed purple colonies, including Enterococcus gallinarum and E. raffinosus, which were later identified using the Vitek card. All VRE isolates were identified by the multiplex PCR method; 100 were vanA-positive E. faecium, 8 were vanA- and vanC-1-positive E. gallinarum, and 1 was vanA-positive E. raffinosus.

CONCLUSIONS

For VRE surveillance, culturing the isolates in chromID VRE after broth enrichment appears to be an accurate, rapid, and easy method for routine screening test. Multiplex PCR is relatively expensive and needs skilled techniques for detecting VRE, but it can be an auxiliary tool for rapid detection of genotype during a VRE outbreak.

摘要

背景

准确且早期检测耐万古霉素肠球菌(VRE)对于控制医院感染至关重要。在本研究中,我们评估了一种选择性显色琼脂培养基和多重聚合酶链反应(PCR)用于检测VRE的效用,并将这两种技术与检测VRE的传统培养方法进行了比较。

方法

我们采用以下3种方法检测粪便标本中的VRE感染:常规培养方法、在选择性显色琼脂培养基(chromID VRE,法国生物梅里埃公司)中培养以及使用Seeplex® VRE ACE检测试剂盒(韩国Seegene公司)并额外进行vanC基因PCR的多重PCR。

结果

通过常规培养方法,我们从100份粪便标本中分离出109株VRE菌株。在chromID VRE中,所有分离株均呈现紫色菌落,包括鹑鸡肠球菌和棉子糖肠球菌,随后使用Vitek卡进行鉴定。所有VRE分离株均通过多重PCR方法鉴定;100株为vanA阳性粪肠球菌,8株为vanA和vanC - 1阳性鹑鸡肠球菌,1株为vanA阳性棉子糖肠球菌。

结论

对于VRE监测,肉汤增菌后在chromID VRE中培养分离株似乎是一种准确、快速且简便的常规筛查方法。多重PCR相对昂贵且检测VRE需要熟练技术,但它可作为VRE暴发期间快速检测基因型的辅助工具。

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