Proudy I, Bouglé D, Coton E, Coton M, Leclercq R, Vergnaud M
Service de Microbiologie, EA 2128 Interactions Hôte et Microorganismes des Epithéliums, CHU Côte de Nacre, Université de Caen Basse-Normandie, Caen Cedex, France.
J Appl Microbiol. 2008 Jan;104(1):26-34. doi: 10.1111/j.1365-2672.2007.03526.x. Epub 2007 Sep 10.
Enterobacter sakazakii is an emerging food-borne pathogen that can cause rare but severe forms of neonatal meningitis, bacteraemia and necrotizing enterocolitis. A rapid typing method at the strain level is needed to determine the monoclonality or polyclonality of the isolates during outbreaks.
The BOX-PCR fingerprinting technique, which targets the repetitive BOX sequences, and sequencing of the flagellin gene, fliC, were evaluated against a panel of 27 Ent. sakazakii strains from clinical and environmental sources. The typeability and discriminatory power of the techniques were compared with those of pulsed-field gel electrophoresis (PFGE), the reference genotyping method. BOX-PCR results yielded 92% agreement with PFGE results, whereas fliC gene sequencing was poorly discriminative.
In our study, BOX-PCR and PFGE were similarly discriminatory to type Ent. sakazakii strains. The weak variability of the Ent. sakazakii fliC gene was related to the absence of the variable central domain present in most fliC genes of Enterobacteriaceae.
The BOX-PCR typing provides an accurate discrimination and a rapid answer to identify clonal isolates of Ent. sakazakii.
阪崎肠杆菌是一种新出现的食源性病原体，可引起罕见但严重的新生儿脑膜炎、菌血症和坏死性小肠结肠炎。在疫情暴发期间，需要一种菌株水平的快速分型方法来确定分离株的单克隆性或多克隆性。
针对来自临床和环境来源的27株阪崎肠杆菌菌株，评估了靶向重复BOX序列的BOX-PCR指纹技术和鞭毛蛋白基因fliC的测序。将这些技术的分型能力和鉴别力与参考基因分型方法脉冲场凝胶电泳（PFGE）进行了比较。BOX-PCR结果与PFGE结果的一致性为92%，而fliC基因测序的鉴别力较差。
在我们的研究中，BOX-PCR和PFGE对阪崎肠杆菌菌株进行分型的鉴别力相似。阪崎肠杆菌fliC基因的弱变异性与大多数肠杆菌科fliC基因中存在的可变中央结构域缺失有关。
BOX-PCR分型为鉴定阪崎肠杆菌的克隆分离株提供了准确的鉴别和快速的答案。
