Rapid and accurate identification of human isolates of Pasteurella and related species by sequencing the sodA gene
Rapid and accurate identification of human isolates of Pasteurella and related species by sequencing the sodA gene
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DOI：
10.1128/jcm.43.5.2307-2314.2005
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发表时间：
2005-05-01
影响因子：
9.4
通讯作者：
Gaillot, O
中科院分区：
文献类型：
--
作者：
Gautier, AL;Dubois, D;Gaillot, O
巴氏杆菌和相关细菌的鉴定仍然是一个挑战。在这里，一个449- 473-bp的片段（sodA（int））内部的sodA基因，编码锰依赖性超氧化物歧化酶，扩增和测序与一对简并引物的模式菌株巴斯德氏菌（18株），鸡杆菌（1株），和曼氏菌（5株）物种。基于sodA（int）的系统发育树与相应16 S rRNA基因序列分析推断的结果基本一致，狭义巴斯德菌群（多杀性巴斯德菌、犬巴斯德菌、达格巴氏巴斯德菌和口巴氏巴斯德菌）的成员形成一个单系群，而鸡杆菌属和曼氏巴斯德菌属是独立的单系属。然而，sodA（int）序列显示出明显高于相应的16 S rRNA基因的分歧，证实sodA是一个有效的目标，以区分相关物种。通过比较其sodA（int）序列与模式种的序列，成功鉴定了33株由参考实验室表型分配为13种巴氏杆菌属的独立人类临床分离株。在这项工作的过程中，我们确定了第一个从人类临床标本中报告的鸡杆菌分离株。临床分离株的sodA（int）序列与相应的模式菌株的sodA（int）序列显示小于2.5%的差异，除了嗜肺巴氏杆菌分离株，其彼此密切相关（> 98% sodA（int）序列同一性），但与模式菌株仅共享92% sodA（int）同一性。当测序设施可用时，本文所述方法为巴斯德菌分离株的种属鉴定提供了快速准确的工具。
The identification of Pasteurella and related bacteria remains a challenge. Here, a 449- to 473-bp fragment (sodA(int)) internal to the sodA gene, encoding the manganese-dependent superoxide dismutase, was amplified and sequenced with a single pair of degenerate primers from the type strains of Pasteurella (18 strains), Gallibacterium (1 strain), and Mannheimia (5 strains) species. The sodA(int)-based phylogenetic tree was in general agreement with that inferred from the analysis of the corresponding 16S rRNA gene sequences, with members of the Pasteurella sensu stricto cluster (Pasteurella multocida, Pasteurella canis, Pasteurella dagmatis, and Pasteurella stomatis) forming a monophyletic group and Gallibacterium and Mannheimia being independent monophyletic genera. However, the sodA(int) sequences showed a markedly higher divergence than the corresponding 16S rRNA genes, confirming that sodA is a potent target to differentiate related species. Thirty-three independent human clinical isolates phenotypically assigned to 13 Pasteurella species by a reference laboratory were successfully identified by comparing their sodA(int) sequences to those of the type species. In the course of this work, we identified the first Gallibacterium anatis isolate ever reported from a human clinical specimen. The sodA(int) sequences of the clinical isolates displayed less than 2.5% divergence from those of the corresponding type strains, except for the Pasteurella pneumotropica isolates, which were closely related to each other (> 98% sodA(int) sequence identity) but shared only 92% sodA(int) identity with the type strain. The method described here provides a rapid and accurate tool for species identification of Pasteurella isolates when access to a sequencing facility is available.