Institut für Medizinische Mikrobiologie und Hygiene, TU Dresden, Fiedlerstrasse 42, D-01307 Dresden, Germany.
J Clin Microbiol. 2011 Dec;49(12):4052-8. doi: 10.1128/JCM.00687-11. Epub 2011 Sep 28.
Sequence-based typing (SBT) is the internationally recognized standard method for genotyping Legionella pneumophila. To date all strains of serogroup 1 (SG1) and some of SGs 2 to 14 yield a seven-allele profile and can be assigned a sequence type (ST). However, for some strains belonging to SGs 2 to 14, the targeted region of the neuA gene could not be amplified using the published standard primers. We determined the DNA sequence of a neuA gene homolog located in the lipopolysaccharide synthesis locus of strain Dallas-1E. By using newly designed degenerate consensus primers based on the neuA homolog in strains Dallas-1E, Philadelphia-1, Paris, Lens, and Corby, we were able to obtain DNA sequences for all 48 non-SG1 strains which were untypeable by the standard method. Our data show that the neuA gene is present in all L. pneumophila strains but differs significantly in some non-SG1 strains at both the DNA and amino acid levels. The new primers can be used to amplify and sequence the neuA gene in all strains and can substitute for the standard primers. This offers the possibility of assigning an ST to all strains of L. pneumophila.
基于序列的分型(SBT)是国际公认的鉴定嗜肺军团菌基因分型的标准方法。迄今为止,所有血清群 1(SG1)菌株和部分血清群 2 至 14 的菌株产生一个七等位基因谱,并可被分配一个序列型(ST)。然而,对于某些属于血清群 2 至 14 的菌株,使用已发表的标准引物无法扩增neuA 基因的目标区域。我们确定了位于达拉斯-1E 菌株脂多糖合成基因座中的 neuA 基因同源物的 DNA 序列。通过使用基于菌株达拉斯-1E、费城-1、巴黎、 Lens 和 Corby 的 neuA 同源物设计的新简并共识引物,我们能够获得所有 48 株无法通过标准方法进行分型的非 SG1 菌株的 DNA 序列。我们的数据表明,neuA 基因存在于所有嗜肺军团菌菌株中,但在一些非 SG1 菌株中,无论是在 DNA 还是氨基酸水平上,都存在显著差异。新引物可用于扩增和测序所有菌株的 neuA 基因,并可替代标准引物。这为所有嗜肺军团菌菌株分配 ST 提供了可能。