Lopes Ana Catarina S, Rodrigues Juliana Falcão, Clementino Maysa B M, Miranda Catia A C, Nascimento Ana Paula A, de Morais Júnior Marcos Antônio
Departamento de Medicina Tropical, Laboratório de Imunopatologia Keizo Asami, Universidade Federal de Pernambuco, Recife, PE, 50732-970, Brazil.
Mem Inst Oswaldo Cruz. 2007 Nov;102(7):827-32. doi: 10.1590/s0074-02762007005000113. Epub 2007 Dec 11.
PCR analysis of 16S-23S internal transcribed spacer (PCR ribotyping) and tRNA intergenic spacer (tDNA-PCR) were evaluated for their effectiveness in identification of clinical strains of Klebsiella pneumoniae and differentiation with related species. For this purpose both methods were applied to forty-three clinical isolates biochemically identified as K. pneumoniae subsp. pneumoniae isolated from patients clinical specimens attended at five hospitals in three Brazilian cities. References strains of K. pneumoniae subsp. pneumoniae, K. pneumoniae subsp. ozaenae, K. oxytoca, K. planticola and Enterobacter aerogenes were also analyzed. Both PCR methods showed specific patterns for each species. A conserved PCR ribotype pattern was observed for all clinical K. pneumoniae isolates, while differing from other related analyzed species. tDNA-PCR revealed five distinct patterns among the K. pneumoniae clinical isolates studied, demonstrating a predominant group with 90.6% of isolates presenting the same pattern of K. pneumoniae type strain. Both PCR-based methods were not able to differentiate K. pneumoniae subspecies. On the basis of the results obtained, both methods were efficient to differentiate the Klebsiella species analyzed, as well as E. aerogenes. Meanwhile tDNA-PCR revealed different tRNA arrangements in K. pneumoniae, suggesting intra-species heterogeneity of their genome organization, the polymorphism of the intergenic spacers between 16S and 23S rRNA genes appears to be highly conserved whithin K. pneumoniae clinical isolates, showing that PCR ribotyping can be an useful tool for identification of K. pneumoniae isolates.
评估了16S - 23S内转录间隔区的PCR分析（PCR核糖体分型）和tRNA基因间间隔区（tDNA - PCR）在鉴定肺炎克雷伯菌临床菌株以及与相关菌种进行区分方面的有效性。为此，将这两种方法应用于从巴西三个城市的五家医院收治患者的临床标本中生化鉴定为肺炎克雷伯菌亚种肺炎克雷伯菌的43株临床分离株。还分析了肺炎克雷伯菌亚种肺炎克雷伯菌、肺炎克雷伯菌亚种臭鼻亚种、产酸克雷伯菌、植生克雷伯菌和产气肠杆菌的参考菌株。两种PCR方法对每个菌种都显示出特定的模式。在所有临床肺炎克雷伯菌分离株中观察到一种保守的PCR核糖体分型模式，与其他相关分析菌种不同。tDNA - PCR在所研究的肺炎克雷伯菌临床分离株中揭示了五种不同的模式，显示出一个主要群体，90.6%的分离株呈现与肺炎克雷伯菌模式菌株相同的模式。两种基于PCR的方法均无法区分肺炎克雷伯菌的亚种。根据获得的结果，两种方法都能有效地区分所分析的克雷伯菌属菌种以及产气肠杆菌。同时，tDNA - PCR揭示了肺炎克雷伯菌中不同的tRNA排列，表明其基因组组织存在种内异质性，16S和23S rRNA基因之间基因间间隔区的多态性在肺炎克雷伯菌临床分离株中似乎高度保守，表明PCR核糖体分型可作为鉴定肺炎克雷伯菌分离株的有用工具。
