Discrimination of Acinetobacter genomic species by AFLP fingerprinting
Discrimination of Acinetobacter genomic species by AFLP fingerprinting
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DOI：
10.1099/00207713-47-4-1179
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发表时间：
1997-10-01
期刊：
影响因子：
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通讯作者：
Dijkshoorn, L
中科院分区：
文献类型：
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作者：
Janssen, P;Maquelin, K;Dijkshoorn, L
AFLP是一种基于限制性内切片段选择性PCR扩增的新型基因组指纹图谱方法。研究了该方法在不动杆菌属基因组物种分化中的适用性。共分析了151个分类菌株（代表18个基因组种，包括类型菌株、参考菌株和野外菌株）和8个未分类菌株。通过使用一组限制性内切酶（HindIII和TaqI）和一组特定的选择性PCR引物，可以将所有菌株分配到正确的基因组种，并将所有组正确分离，最小种内相似性水平为29 ~ 74%。菌株属于基因组种8（严格lwoffii不动杆菌）和9聚在一个簇中。密切相关的DNA类群1（钙酸不动杆菌）、2（鲍曼不动杆菌）、3和13TU （sensu Tjernberg & Ursing 1989）可明显区分，种内连锁水平超过50%。独立描述的基因组物种13BJ （sensu Bouvet & Jeanjean 1989）和14TU的联系度相对较低（33%）。尽管先前的DNA-DNA杂交研究似乎证明了这些基因组物种的统一，但AFLP分析实际上将13BJ-14TU群划分为三个分离良好的亚群。最后，从不同来源和起源获得的4个未分类菌株令人信服地分组在一起，相似连锁水平约为50%。这些菌株与其他155株菌株的AFLP模式没有相似之处，可能代表了迄今未被描述的不动杆菌物种。基于这些结果，AFLP可以作为基因组物种描述的重要辅助方法。此外。由于AFLP提供了对不动杆菌分类群的种下结构的详细了解，该方法也代表了在不动杆菌流行病学中确认菌株身份的高效手段。
AFLP is a novel genomic fingerprinting method based on the selective PCR amplification of restriction fragments. The usability of this method for the differentiation of genomic species in the genus Acinetobacter was investigated. A total of 151 classified strains (representing 18 genomic species, including type, reference, and field strains) and 8 unclassified strains were analyzed. By using a single set of restriction enzymes (HindIII and TaqI) and one particular set of selective PCR primers, all strains could be allocated to the correct genomic species and all groups were properly separated, with minimal intraspecific similarity levels ranging from 29 to 74%. Strains belonging to genomic species 8 (Acinetobacter lwoffii sensu stricto) and 9 grouped together in one cluster. The closely related DNA groups 1 (Acinetobacter calcoaceticus), 2 (Acinetobacter baumannii), 3, and 13TU (sensu Tjernberg & Ursing 1989) were clearly distinguishable, with intraspecific linkage levels above 50%. Strains of the independently described genomic species 13BJ (sensu Bouvet & Jeanjean 1989) and 14TU linked together at a relatively low level (33%). Although a previous DNA-DNA hybridization study seemed to justify the unification of these genomic species, AFLP analysis actually divides the 13BJ-14TU group into three well-separated subgroups. Finally, four unclassified strains obtained from diverse sources and origins grouped convincingly together, with a similarity linkage level of approximately 50%. These strains showed no similarities in their AFLP patterns with any of the other 155 strains studied and may represent a thus-far-undescribed Acinetobacter species. Based on these results, AFLP should be regarded as an important auxiliary method for the delineation of genomic species. Furthermore. because AFLP provides a detailed insight into the infraspecific structure of Acinetobacter taxa, the method also represents a highly effective means for the confirmation of strain identity in the epidemiology of acinetobacters.