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Species-Specific Identification of Campylobacters by Partial 16S rRNA Gene Sequencing

机译:通过部分16S rRNA基因测序对弯曲菌进行物种特异性鉴定

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摘要

Species-specific identification of campylobacters is problematic, primarily due to the absence of suitable biochemical assays and the existence of atypical strains. 16S rRNA gene (16S rDNA)-based identification of bacteria offers a possible alternative when phenotypic tests fail. Therefore, we evaluated the reliability of 16S rDNA sequencing for the species-specific identification of campylobacters. Sequence analyses were performed by using almost 94% of the complete 16S rRNA genes of 135 phenotypically characterized Campylobacter strains, including all known taxa of this genus. It was shown that 16S rDNA analysis enables specific identification of most Campylobacter species. The exception was a lack of discrimination among the taxa Campylobacter jejuni and C. coli and atypical C. lari strains, which shared identical or nearly identical 16S rDNA sequences. Subsequently, it was investigated whether partial 16S rDNA sequences are sufficient to determine species identity. Sequence alignments led to the identification of four 16S rDNA regions with high degrees of interspecies variation but with highly conserved sequence patterns within the respective species. A simple protocol based on the analysis of these sequence patterns was developed, which enabled the unambiguous identification of the majority of Campylobacter species. We recommend 16S rDNA sequence analysis as an effective, rapid procedure for the specific identification of campylobacters.
机译:弯曲菌的物种特异性鉴定是有问题的,这主要是由于缺乏合适的生化分析和非典型菌株的存在。当表型测试失败时,基于16S rRNA基因(16S rDNA)的细菌鉴定提供了一种可能的替代方法。因此,我们评估了16S rDNA测序用于弯曲菌物种特异性鉴定的可靠性。序列分析是通过使用135个具有表型特征的弯曲杆菌菌株(包括该属的所有已知分类单元)的几乎全部16S rRNA基因的94%进行的。结果表明,16S rDNA分析能够特异性鉴定大多数弯曲杆菌。例外是空肠弯曲杆菌,大肠弯曲杆菌和非典型弯曲弧菌菌株之间缺乏区分,它们共享相同或几乎相同的16S rDNA序列。随后,研究了部分16S rDNA序列是否足以确定物种同一性。序列比对导致鉴定出具有高度种间变异性但在各个物种内具有高度保守的序列模式的四个16S rDNA区域。基于这些序列模式的分析,开发了一种简单的协议,可以明确鉴定大多数弯曲杆菌属。我们建议使用16S rDNA序列分析作为快速,准确鉴定弯曲杆菌的有效方法。

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