Université de Lyon, France Research Group on 'Bacterial Opportunistic Pathogens and Environment', UMR 5557 Ecologie Microbienne, CNRS, VetAgro Sup and Université Lyon 1, 43 Boulevard du 11 Novembre 1918, 69622 Villeurbanne Cedex, France.
J Appl Microbiol. 2011 Nov;111(5):1185-93. doi: 10.1111/j.1365-2672.2011.05120.x. Epub 2011 Aug 30.
Aim of the study is to identify accurately Stenotrophomonas maltophilia isolates recovered from environmental and clinical samples.
Recovery of Sten. maltophilia-like isolates from soil samples using the vancomycin, imipenem, amphotericin B (VIA) selective agar medium enabled distinction of various morphotype colonies. A set of soil and clinical isolates was tested for species identification using different methods. 16S rDNA analyses showed the dark green with a blue halo morphotype to be typical Sten. maltophilia strains. The API-20NE, Vitek-2 and Biolog phenotypic analyses typically used for the identification of clinical isolates did not perform well on these soil isolates. The species-specific PCR screening targeting Sten. maltophilia 23S rDNA and the multiplex smeD/ggpS PCR, differentiating Sten. maltophilia from Stenotrophomonas rhizophila, were tested for improvement of these identification schemes. The latter multiplex PCR identified all isolates tested in this study, whatever be their origin.
Isolation on VIA medium and confirmation of Sten. maltophilia species membership by smeD PCR is proposed to identify environmental and clinical isolates of Sten. maltophilia.
The proposed approach enables isolation and identification of Sten. maltophilia from different environments in an easy and rapid way. This approach will be useful to accurately manage studies on the abundance and distribution of Sten. maltophilia in hospital and nonhospital environments.
本研究旨在准确鉴定从环境和临床样本中分离出的嗜麦芽寡养单胞菌。
使用万古霉素、亚胺培南、两性霉素 B(VIA)选择性琼脂培养基从土壤样本中回收嗜麦芽寡养单胞菌样分离物,可区分不同形态的菌落。使用不同方法对一组土壤和临床分离物进行了物种鉴定测试。16S rDNA 分析表明,深绿色带蓝色晕环的形态为典型的嗜麦芽寡养单胞菌株。API-20NE、Vitek-2 和 Biolog 表型分析通常用于临床分离物的鉴定,但对这些土壤分离物的效果不佳。针对嗜麦芽寡养单胞菌 23S rDNA 的特异性 PCR 筛选和区分嗜麦芽寡养单胞菌和嗜麦芽寡养单胞菌的多重 smeD/ggpS PCR 用于改进这些鉴定方案。后者的多重 PCR 鉴定了本研究中测试的所有分离物,无论其来源如何。
建议在 VIA 培养基上进行分离,并通过 smeD PCR 确认嗜麦芽寡养单胞菌的物种归属,以鉴定环境和临床分离物中的嗜麦芽寡养单胞菌。
所提出的方法可方便、快速地从不同环境中分离和鉴定嗜麦芽寡养单胞菌。这种方法将有助于准确管理嗜麦芽寡养单胞菌在医院和非医院环境中的丰度和分布研究。