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通过多态性DNA的随机扩增或DNA大片段限制性酶切片段的脉冲场凝胶电泳对铜绿假单胞菌进行比较分型。

Comparative typing of Pseudomonas aeruginosa by random amplification of polymorphic DNA or pulsed-field gel electrophoresis of DNA macrorestriction fragments.

作者信息

Renders N, Römling U, Verbrugh H, van Belkum A

机构信息

Department of Bacteriology, University Hospital Rotterdam, The Netherlands.

出版信息

J Clin Microbiol. 1996 Dec;34(12):3190-5. doi: 10.1128/jcm.34.12.3190-3195.1996.

Abstract

Eighty-seven strains of Pseudomonas aeruginosa were typed by random amplification of polymorphic DNA (RAPD) and pulsed-field gel electrophoresis (PFGE) of macrorestriction fragments. Stains were clustered on the basis of interpretative criteria as presented previously for the PFGE analysis. Clusters of strains were also defined on the basis of epidemiological data and subsequently reanalyzed by RAPD. It was found that in an RAPD assay employing the enterobacterial repetitive intergenic consensus sequence ERIC2 as a primer, single band differences can be ignored; in this case, clonally related strains could be grouped as effectively and reliably as with PFGE. These data could be corroborated by the use of other primer species. However, some primers either showed reduced resolution or, in contrast, identified DNA polymorphisms beyond epidemiologically and PFGE-defined limits. Apparently, different primers define different windows of genetic variation. It is suggested that criteria for interpretation of the ERIC2 PCR fingerprints can be simple and straightforward: when single band differences are ignored, RAPD-determined grouping of P. aeruginosa is congruent with that obtained by PFGE. Consequently, this implies that RAPD can be used with trust as a first screen in epidemiological characterization of P. aeruginosa. The ability to measure the rate of molecular evolution of the P. aeruginosa genome clearly depends on the choice of restriction enzyme or primer when RAPD or PFGE, respectively, is applied for the detection of DNA polymorphisms.

摘要

通过随机扩增多态性DNA(RAPD)和宏限制性片段的脉冲场凝胶电泳(PFGE)对87株铜绿假单胞菌进行分型。菌株根据先前用于PFGE分析的解释标准进行聚类。还根据流行病学数据定义菌株簇,随后通过RAPD重新分析。发现在使用肠杆菌重复基因间共有序列ERIC2作为引物的RAPD分析中,单带差异可以忽略;在这种情况下,克隆相关菌株可以与PFGE一样有效且可靠地分组。使用其他引物种类可以证实这些数据。然而,一些引物要么分辨率降低,要么相反,识别出超出流行病学和PFGE定义范围的DNA多态性。显然,不同的引物定义了不同的遗传变异窗口。建议对ERIC-PCR指纹图谱的解释标准可以简单直接:当忽略单带差异时,RAPD确定的铜绿假单胞菌分组与PFGE获得的分组一致。因此,这意味着RAPD可以放心地用作铜绿假单胞菌流行病学特征分析的初步筛选方法。当分别应用RAPD或PFGE检测DNA多态性时,测量铜绿假单胞菌基因组分子进化速率的能力显然取决于限制性内切酶或引物的选择。

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