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The development of a 16S rRNA gene based PCR for the identification of Streptococcus pneumoniae and comparison with four other species specific PCR assays

机译:开发基于16s rRNa基因的pCR鉴定肺炎链球菌并与其他4种特异性pCR检测方法进行比较

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摘要

Background: Streptococcus pneumoniae is one of the most frequently encountered pathogens in humans but its differentiation from closely related but less pathogenic streptococci remains a challenge.Methods: This report describes a newly-developed PCR assay (Spne-PCR), amplifying a 217 bp product of the 16S rRNA gene of S. pneumoniae, and its performance compared to other genotypic and phenotypic tests.Results: The new PCR assay designed in this study, proved to be specific at 57 degrees C for S. pneumoniae, not amplifying S. pseudopneumoniae or any other streptococcal strain or any strains from other upper airway pathogenic species. PCR assays (psaA, LytA, ply, spn9802-PCR) were previously described for the specific amplification of S. pneumoniae, but psaA-PCR was the only one found not to cross-react with S. pseudopneumoniae.Conclusion: Spne-PCR, developed for this study, and psaA-PCR were the only two assays which did not mis-identify S. pseudopneumoniae as S. pneumoniae. Four other PCR assays and the AccuProbe assay were unable to distinguish between these species.
机译:背景:肺炎链球菌是人类中最常见的病原体之一,但如何与密切相关但病原性较低的链球菌区分开仍然是一个挑战。肺炎链球菌的16S rRNA基因的克隆及其与其他基因型和表型试验的性能比较。或任何其他链球菌菌株或来自其他上呼吸道致病菌种的任何菌株。先前曾描述过PCR检测(psaA,LytA,ply,spn9802-PCR)特异性扩增肺炎链球菌的方法,但psaA-PCR是唯一发现不与假性肺炎链球菌交叉反应的方法。为这项研究开发了psaA-PCR,这是唯一没有将假性肺炎链球菌误认为肺炎链球菌的两种检测方法。其他四种PCR检测和AccuProbe检测无法区分这些物种。

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