rpoB gene sequence-based identification of Staphylococcus species
rpoB gene sequence-based identification of Staphylococcus species
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DOI：
10.1128/jcm.40.4.1333-1338.2002
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发表时间：
2002-04-01
影响因子：
9.4
通讯作者：
Raoult, D
中科院分区：
文献类型：
--
作者：
Drancourt, M;Raoult, D
对糖溶葡萄球菌、鲁顿葡萄球菌、S冠状葡萄球菌和中间葡萄球菌的rpoB基因进行了全序列测定，并对另外27种葡萄球菌进行了部分序列测定。完整的rpoB序列长度在3,452-3,845bp之间，GC含量为36.8-39.2%。部分序列种间同源性为71.6%~93.6%，种内差异为0.08%~0.8%。除了少数例外，从部分rpoB序列推断的系统发育关系与以前通过DNA-DNA杂交研究和16S核糖体DNA基因序列和部分HSP60基因序列分析得出的结果一致。我们建立的葡萄球菌rpoB序列数据库使我们能够发展一种分子方法来鉴定葡萄球菌分离株，然后对751个碱基的扩增片段进行直接测序。盲法对10株葡萄球菌进行正确鉴定，对10株革兰氏阳性和革兰氏阴性非葡萄球菌均未检测出阳性结果。我们建议将rpoB基因的部分序列作为准确鉴定葡萄球菌分离株的新工具。
The complete sequence of rpoB, the gene encoding the beta subunit of RNA polymerase was determined for Staphylococcus saccharolyticus, Staphylococcus lugdunensis, S taphylococcus caprae, and Staphylococcus intermedius and partial sequences were obtained for an additional 27 Staphylococcus species. The complete rpoB sequences varied in length from 3,452 to 3,845 bp and had a 36.8 to 39.2% GC content. The partial sequences had 71.6 to 93.6% interspecies homology and exhibited a 0.08 to 0.8% intraspecific divergence. With a few exceptions, the phylogenetic relationships inferred from the partial rpoB sequences were in agreement with those previously derived from DNA-DNA hybridization studies and analyses of 16S ribosomal DNA gene sequences and partial HSP60 gene sequences. The staphylococcal rpoB sequence database we established enabled us to develop a molecular method for identifying Staphylococcus isolates by PCR followed by direct sequencing of the 751-bp amplicon. In blind tests, this method correctly identified 10 Staphylococcus isolates, and no positive results were obtained with 10 non-Staphylococcus gram-positive and gram-negative bacterial isolates. We propose partial sequencing of the rpoB gene as a new tool for the accurate identification of Staphylococcus isolates.