A specific polymerase chain reaction test for the identification of Streptococcus pneumoniae
A specific polymerase chain reaction test for the identification of Streptococcus pneumoniae
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DOI：
10.1016/j.diagmicrobio.2011.02.005
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发表时间：
2011-05-01
影响因子：
2.9
通讯作者：
Fayet, Olivier A.
中科院分区：
文献类型：
--
作者：
Prere, Marie-Francoise;Fayet, Olivier A.
通过比较口腔链球菌和肺炎链球菌的随机引物聚合酶链式反应(PCR)基因组图谱，我们确定了一条434bp的基因组片段，该片段是肺炎链球菌所特有的。根据这一常见片段的核苷酸序列，设计了一对引物，并在一组包括主要链球菌的菌株上进行了测试。其中一种叫狭小链球菌，其扩增产物与肺炎链球菌的长度相同。对链球菌和肺炎链球菌扩增片段的序列比较发现了几个变异，这些变异被用来定义一组新的引物，给出了181个碱基的肺炎链球菌特异片段。该扩增片段包含一个编码糖专一性渗透酶的基因的5‘末端部分和一个基因间序列。对与临床分离相对应的257株侵袭性肺炎链球菌和153株非肺炎口腔链球菌进行了PCR检测，并对3株假肺炎链球菌进行了检测。用这些引物只能对肺炎链球菌进行扩增。此外，该测试还成功地对10株与肺炎链球菌疾病有关的非典型肺炎链球菌进行了评估。因此，在这项研究中，我们建立了一种简单的聚合酶链式反应试验来区分肺炎链球菌和其他链球菌(包括假肺炎链球菌)的能力，从而使快速和准确的诊断成为可能。(C)2011 Elsevier Inc.保留所有权利。
Using an approach based on the comparison of arbitrary primer polymerase chain reaction (PCR) genomic profiles from oral streptococci and Streptococcus pneumoniae strains, we identified a 434-bp genomic fragment apparently specific for S. pneumoniae. From the nucleotidic sequence of this common fragment, a pair of primers was designed and tested on a set of strains comprising the major Streptococcus species. One species, S. anginosus, gave an amplification product of the same length as S. pneumoniae. Sequence comparison of the S. anginosus and S. pneumoniae amplicons revealed several variations which were used to define a new set of primers giving a 181-bp S. pneumoniae specific fragment. The amplified fragment contains the 5' terminal part of a gene encoding a putative sugar-specific permease and an intergenic sequence. The PCR test was evaluated on 257 strains of invasive S. pneumoniae corresponding to clinical isolates and on 153 non-pneumoniae oral streptococci strains; in addition, 3 S. pseudopneumoniae strains were tested. With these primers, an amplification product was only obtained with the S. pneumoniae strains. Moreover, the test was successfully evaluated on 10 atypical S. pneumoniae strains related to pneumococcal diseases. In this study, we therefore established the capacity of a simple PCR test to discriminate S. pneumoniae from other Streptococci (including S. pseudopneumoniae), thus allowing rapid and accurate diagnosis. (C) 2011 Elsevier Inc. All rights reserved.