PCR-amplified 16S and 23S rDNA restriction analysis for the identification of Acinetobacter strains at the DNA group level
PCR-amplified 16S and 23S rDNA restriction analysis for the identification of Acinetobacter strains at the DNA group level
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DOI：
10.1016/s0923-2508(97)82453-9
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发表时间：
1997-12-01
影响因子：
2.6
通讯作者：
Ibrahim, A
中科院分区：
文献类型：
--
作者：
García-Arata, MI;Gerner-Smidt, P;Ibrahim, A
不动杆菌属在表型上是相当同质的，但在基因型上是异质的。在这项研究中，一个简单的方法的基础上限制性分析的PCR扩增的大片段（4.5 kb）的大多数核糖体操纵子（16 S和23 S核糖体基因和间隔之间）进行了研究。67个收集菌株属于20个DNA组提出，直到1993年进行了研究。使用酶Sau 3AI，获得25个DNA图谱。属于DNA组1、3、6、TU 13和TU 15的菌株各显示两个谱，DNA组4、5和7显示具有变体的谱，所述变体显示不太密集的额外条带。其余12组显示出12种不同的特征。所获得的配置文件是DNA组特异性的，除了一个配置文件之间共享的未命名的DNA组3和很少遇到的基因型相关的DNA组。这两个DNA组可以通过使用酶Hinf 1分离。以双盲方式选择先前通过标准DNA-DNA杂交表征的25个额外临床分离株，用于在DNA组水平进行鉴定，以检查测定的可靠性。PCR扩增16 S和23 S rDNA酶切分析是一种准确、快速的DNA组水平鉴定不动杆菌属的方法。
The genus Acinetobacter is phenotypically rather homogeneous, but genotypically heterogeneous. In this study, a simple method based on restriction analysis of a PCR-amplified large fragment (4.5 kb) of most of the ribosomal operon (16S and 23S ribosomal genes and the spacer in-between) was investigated. Sixty-seven collection strains belonging to the 20 DNA groups proposed until 1993 were studied. Using the enzyme Sau3AI, 25 DNA profiles were obtained. Strains belonging to DNA groups 1, 3, 6, TU13 and TU15 showed two profiles each, and DNA groups 4, 5 and 7 showed profiles with variants showing less intensive additional bands. The remaining 12 groups showed 12 different profiles. The profiles obtained were DNA-group-specific except for one profile which was shared between the unnamed DNA group 3 and a rarely encountered genotypically related DNA group. These two DNA groups could be separated by using the enzyme Hinf1. Twenty-five additional clinical isolates previously characterized by standard DNA-DNA hybridization were selected in a double-blind fashion for identification at the DNA group level to check the reliability of the assay. All strains were correctly identified at the DNA group level, PCR-amplified 16S and 23S rDNA restriction analysis is both an accurate and rapid method for the identification of Acinetobacter at the DNA group level.