Maternal oral origin of Fusobacterium nucleatum in adverse pregnancy outcomes as determined using the 16S-23S rRNA gene intergenic transcribed spacer region
Maternal oral origin of Fusobacterium nucleatum in adverse pregnancy outcomes as determined using the 16S-23S rRNA gene intergenic transcribed spacer region
复制标题
DOI：
10.1099/jmm.0.049452-0
复制
发表时间：
2013-01-01
影响因子：
3
通讯作者：
Allaker, Robert P.
中科院分区：
文献类型：
--
作者：
Gonzales-Marin, Cecilia;Spratt, David A.;Allaker, Robert P.
具核梭杆菌是一种常见的革兰氏阴性厌氧菌，在口腔中流行，具有定殖羊膜腔和胎儿的能力。然而，F.核质也可能是阴道微生物群的一部分，从那里它可以到达羊膜组织。由于F.有核细胞由五个亚种组成，需要在亚种/菌株水平上进行分析，以确定其确切来源。本研究的目的是：（1）评价16 S-23 SrRNA基因间转录间隔区（ITS）作为区分F.设计一种基于ITS的简化技术来确定F.与不良妊娠结局相关的核质菌株。对应于F. 5个亚种的16 S-23 S rRNA基因ITS区的扩增片段。核质进行克隆和测序，以表征亚种的不同核糖体操纵子。鉴别了可能用于鉴定亚种/菌株的独特长度和序列模式。这些被用来评估F.通过与相应的口腔和阴道母体样品进行序列比较，在新生儿胃吸出物（吞咽的羊水）中鉴定出有核质。随后开发了一种在更敏感的巢式PCR中使用菌株特异性引物的简化技术来分析10对新生儿母体样本。分析ITS区的可变片段可以鉴定F.具核壳亚种来自口腔来源的多形藻可能参与新生儿感染。利用菌株特异性引物，将F。具核壳亚种在患有局部牙周袋的母亲的早产病例中，在新生儿胃吸出物和母亲口腔样本中均检测到多形菌菌株。有趣的是，在所调查的任何病例的阴道样本中均不存在相同的菌株。16 S-23 SrRNA基因ITS区可作为核镰孢菌起源的一个有用工具。本研究的结果有力地表明，F。具核壳亚种口腔来源的多形性可能与妊娠并发症有关。
Fusobacterium nucleatum, a common Gram-negative anaerobe prevalent in the oral cavity, possesses the ability to colonize the amniotic cavity and the fetus. However, F. nucleatum may also be part of the vaginal microbiota from where it could reach the amniotic tissues. Due to the heterogeneity of F. nucleatum, consisting of five subspecies, analysis at the subspecies/strain level is desirable to determine its precise origin. The aims of this study were: (i) to evaluate the use of the 16S-23S rRNA gene intergenic transcribed spacer (ITS) region as a tool to differentiate subspecies of F. nucleatum, and (ii) to design a simplified technique based on the ITS to determine the origin of F. nucleatum strains associated with adverse pregnancy outcomes. Amplified fragments of the 16S-23S rRNA gene ITS region corresponding to the five subspecies of F. nucleatum were subjected to cloning and sequencing to characterize the different ribosomal operons of the subspecies. Distinctive length and sequence patterns with potential to be used for identification of the subspecies/strain were identified. These were used to evaluate the origin of F. nucleatum identified in neonatal gastric aspirates (swallowed amniotic fluid) by sequence comparisons with the respective oral and vaginal maternal samples. A simplified technique using a strain-specific primer in a more sensitive nested PCR was subsequently developed to analyse ten paired neonatal maternal samples. Analysing the variable fragment of the ITS region allowed the identification of F. nucleatum subsp. polymorphum from an oral origin as potentially being involved in neonatal infections. Using a strain-specific primer, the F. nucleatum subsp. polymorphum strain was detected in both neonatal gastric aspirates and maternal oral samples in cases of preterm birth from mothers presenting with localized periodontal pockets. Interestingly, the same strain was not present in the vaginal sample of any case investigated. The 16S-23S rRNA gene ITS can be a useful tool to determine the origin of F nucleatum. The results of this study strongly indicate that F. nucleatum subsp. polymorphum of oral origin could be involved with pregnancy complications.