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首页> 外文期刊>Journal of Clinical Microbiology >Identification of Abiotrophia adiacens and Abiotrophia defectiva by 16S rRNA gene PCR and restriction fragment length polymorphism analysis.
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Identification of Abiotrophia adiacens and Abiotrophia defectiva by 16S rRNA gene PCR and restriction fragment length polymorphism analysis.

机译：通过16S rRNA基因PCR和限制性酶切片段长度多态性分析鉴定双歧杆菌和缺陷型双歧杆菌。

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Abiotrophia adiacens and Abiotrophia defectiva, previously referred to as nutritionally variant streptococci, Streptococcus adjacens and Streptococcus defectivus, respectively, are causes of infective endocarditis. We describe a method of identifying these two species and also of distinguishing them from 15 other major etiological pathogens of infective endocarditis by means of 16S rRNA gene PCR amplification followed by restriction fragment length polymorphism analysis (PCR-RFLP). The 16S rRNA genes were successfully amplified with a set of universal primers from all 17 species of bacteria examined, including viridans group streptococci. The RFLP patterns of A. adiacens and A. defectiva obtained by HaeIII or MspI digestion were readily distinguished from each other and from those of other bacteria. When PCR analysis was performed with the supernatant of a suspension of a boiled colony, the 16S rRNA genes of 80 of 82 isolates (97%) of A. adiacens and all isolates (11 of 11) of A. defectiva were amplified. The HaeIII RFLP patterns of the isolates were the same as those of the corresponding type strains, although 28% of A. adiacens isolates revealed intraspecies polymorphism. The detection limit of this method was 0.1 pg of genomic DNA, as assessed by using the digoxigenin-labeling DNA detection system. Thus, the PCR-RFLP analysis that we developed is applicable for the routine detection of Abiotrophia from clinical specimens.
机译：先前被称为营养变异型链球菌，邻链球菌和缺陷链球菌的重金属厌氧菌和缺氧厌氧菌是引起感染性心内膜炎的原因。我们描述了一种识别这两种物种的方法，还通过16S rRNA基因PCR扩增，然后进行限制性片段长度多态性分析（PCR-RFLP），将它们与传染性心内膜炎的其他15种主要病原体区分开。 16S rRNA基因已成功从一组包括细菌纲链球菌在内的所有17种细菌的通用引物中扩增出来。通过HaeIII或MspI消化获得的拟南芥和缺陷曲霉的RFLP模式很容易彼此区分开，也易于与其他细菌区分。当用煮沸的菌落的悬液的上清液进行PCR分析时，扩增了82个分离的拟南曲霉中的80个（97％）和所有缺陷的拟南曲霉（11个中的11个）的16S rRNA基因。分离株的HaeIII RFLP模式与相应菌株的相同，尽管28％的A. adiacens分离株显示出种内多态性。通过使用洋地黄毒苷标记的DNA检测系统评估，该方法的检测限为0.1 pg基因组DNA。因此，我们开发的PCR-RFLP分析适用于常规从临床标本中检测出Atrotrophia。

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