Development of a recA gene-based identification approach for the the entire Burkholderia genus
Development of a recA gene-based identification approach for the the entire Burkholderia genus
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DOI：
10.1128/aem.71.7.3917-3927.2005
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发表时间：
2005-07-01
影响因子：
4.4
通讯作者：
Mahenthiralingam, E
中科院分区：
文献类型：
--
作者：
Payne, GW;Vandamme, P;Mahenthiralingam, E
伯克霍尔德氏菌属是一个重要的细菌属，包含具有生态、生物技术和病原学意义的菌种。由于分类学的不断修订和几个物种的表型相似性，伯克霍尔德氏菌的正确鉴定是困难的。基于recA基因的遗传方案大大提高了洋葱伯克霍尔德菌复合种的鉴定。然而，为后一种方法开发的PCR受到其对复合物的特异性的限制。通过比对现有的和新的伯克霍尔德氏菌recA序列，我们设计了新的PCR引物，并通过测试一组代表性的伯克霍尔德氏菌菌株来评估其特异性。PCR，然后限制性片段长度多态性分析的869 bp的伯克霍尔德氏菌recA基因的一部分是没有足够的歧视。对recA片段进行核苷酸测序，然后进行系统发育分析，区分了推定的和已知的伯克霍尔德菌属物种以及B的所有成员。洋葱复合体此外，它使伯克霍尔德氏菌属特异性recA PCR的设计，产生一个385 bp的扩增子，其序列也能够区分所有物种的检查。188个新recA基因的系统发育分析，使澄清几个重要的伯克霍尔德菌菌株的分类位置，并揭示了四个新的B的存在。洋葱复合体recA谱系。虽然遗传学不能作为区分B的手段。从临床感染中恢复的洋葱复合体菌株与自然环境中恢复的洋葱复合体菌株相比，它确实有助于鉴定B的克隆菌株类型。洋葱B. cepacia，B. stabilis和B.能够在两个壁龛中居住的ambifaria。
Burkholderia is an important bacterial genus containing species of ecological, biotechnological, and pathogenic interest. With their taxonomy undergoing constant revision and the phenotypic similarity of several species, correct identification of Burkholderia is difficult. A genetic scheme based on the recA gene has greatly enhanced the identification of Burkholderia cepacia complex species. However, the PCR developed for the latter approach was limited by its specificity for the complex. By alignment of existing and novel Burkholderia recA sequences, we designed new PCR primers and evaluated their specificity by testing a representative panel of Burkholderia strains. PCR followed by restriction fragment length polymorphism analysis of an 869-bp portion of the Burkholderia recA gene was not sufficiently discriminatory. Nucleotide sequencing followed by phylogenetic analysis of this recA fragment differentiated both putative and known Burkholderia species and all members of the B. cepacia complex. In addition, it enabled the design of a Burkholderia genus-specific recA PCR that produced a 385-bp amplicon, the sequence of which was also able to discriminate all species examined. Phylogenetic analysis of 188 novel recA genes enabled clarification of the taxonomic position of several important Burkholderia strains and revealed the presence of four novel B. cepacia complex recA lineages. Although the recA phylogeny could not be used as a means to differentiate B. cepacia complex strains recovered from clinical infection versus the natural environment, it did facilitate the identification of clonal strain types of B. cepacia, B. stabilis, and B. ambifaria capable of residing in both niches.