Validation of partial rpoB gene sequence analysis for the identification of clinically important and emerging Acinetobacter species
Validation of partial rpoB gene sequence analysis for the identification of clinically important and emerging Acinetobacter species
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DOI：
10.1099/mic.0.026054-0
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发表时间：
2009-07-01
期刊：
影响因子：
2.8
通讯作者：
La Scola, Bernard
中科院分区：
文献类型：
--
作者：
Gundi, Vijay A. K. B.;Dijkshoorn, Lenie;La Scola, Bernard
属于不动杆菌属的细菌在土壤和水中普遍存在。只有少数物种，包括鲍氏不动杆菌和未命名的不动杆菌基因组物种（gen. sp.）3和13 TU与土壤微生物乙酸钙不动杆菌（Acinetobacter calcoaceticus）结合在A.醋酸钙A.鲍曼不动杆菌（Acb）复合物是重要的医院感染原。大多数物种的生态学、流行病学和病理学尚未完全确定。缺乏实用和准确的方法限制了临床分离株的常规鉴定，从而阻碍了Acb复合菌和其他可能具有临床意义的不动杆菌属的精确鉴定。我们以前确定了一个350 bp的rpoB基因的高度可变区，这似乎是一个有前途的快速分子鉴定的目标。在本研究中，我们验证了这种方法的准确性，对一个集合的参考菌株属于A。醋酸钙杆菌（5株）、不动杆菌属（Acinetobacter gen.sp.3）（29株）、醋酸钙不动杆菌（A. gen.sp.13TU（118株）、A.鲍曼不动杆菌（30株）和A. radioresistens，A. gen. sp. 15TU，A. gen.sp.10、黑腹甲A. gen.sp.11、黑腹甲A. gen. sp. '1 ~ 3'和A. 14TU = 13BJ。这代表了迄今为止最大的分析，比较了大量已充分鉴定的Acb复合体菌株，以评估该复合体内的种内和种间变异。基于部分rpoB序列分析，所有这些都以98.9-100%的种内相关性被正确鉴定。然后，我们应用该工具，以确定99个不动杆菌临床分离株从四个公立医院在法国马赛。所有分离物可以很容易地被鉴定为种，因为它们被分成13种序列类型，与它们各自的模式菌株的序列变异为0-2.6%。99株中有10株为A.溶血性弧菌52株。鲍曼不动杆菌27株。gen.sp.3，5株为A. schindleri，A. Iwoffii和1个A. gen.sp.13TU.其中三个暂时被确定为A。gen. sp. 9.这是首次使用rpoB序列分析在种水平上鉴定一组临床不动杆菌分离株的所有标本。我们的数据强调了A. schindleri作为一种新出现的不动杆菌相关感染的原因，并确认A. gen.sp.3是继A.鲍曼不动杆菌
Bacteria belonging to the genus Acinetobacter are ubiquitous in soil and water. Only a few species, including Acinetobacter baumannii, and the unnamed Acinetobacter genomic species (gen. sp.) 3 and 13TU, which together with the soil organism Acinetobacter calcoaceticus are combined in the A. calcoaceticus-A. baumannii (Acb) complex, have been recognized as important nosocomial infectious agents. The ecology, epidemiology and pathology of most species are not yet well established. Lack of practical and accurate methods limits routine identification of clinical isolates and thus hampers precise identification of those of the Acb complex and other Acinetobacter species of possible clinical significance. We previously identified a 350 bp highly variable zone on the rpoB gene which appeared to be a promising target for rapid molecular identification. In the present study, we validated this method for accuracy on a collection of reference strains belonging to A. calcoaceticus (5 strains), Acinetobacter gen. sp. 3 (29 strains), A. gen. sp. 13TU (118 strains), A. baumannii (30 strains) and one strain each of A. radioresistens, A. gen. sp. 15TU, A. gen. sp. 10, A. gen. sp. 11, A. gen. sp. 'between 1 and 3' and A. gen. sp. 14TU = 13BJ. This represents the largest analysis to date that compares a large number of well-identified strains of the Acb complex to assess the intra- and interspecies, variation within this complex. All were correctly identified with 98.9-100% intraspecies relatedness based on partial rpoB sequence analysis. We then applied this tool to identify 99 Acinetobacter clinical isolates from four public hospitals in Marseille, France. All isolates could easily be identified to species as they were separated into 13 species sequence types with a sequence variance of 0-2.6% from their respective type strains. Of these 99 isolates, 10 were A. haemolyticus, 52 were A. baumannii, 27 were A. gen. sp. 3, 5 were A. schindleri, 1 was A. Iwoffii, and 1 was A. gen. sp. 13TU. Three were provisionally identified as A. gen. sp. 9. This is the first work to identify all specimens of a set of clinical Acinetobacter isolates at species level using rpoB sequence analysis. Our data emphasize the recognition of A. schindleri as an emerging cause of Acinetobacter-related infection and confirm that A. gen. sp. 3 is the second most commonly isolated Acinetobacter species after A. baumannii in patients.