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Fungal rDNA signatures in coronary atherosclerotic plaques

机译:冠状动脉粥样斑块中的真菌rDNA签名

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Bacterial DNA has been found in coronary plaques and it has therefore been concluded that bacteria may play a role as trigger factors in the chronic inflammatory process underlying coronary atherosclerosis. However, the microbial spectrum is complex and it is not known whether microorganisms other than bacteria are involved in coronary disease. Fungal 18S rDNA signatures were systematically investigated in atherosclerotic tissue obtained through catheter-based atherectomy of 38 patients and controls (unaffected coronary arteries) using clone libraries, denaturating gradient gel analysis (DGGE), in situ hybridization and fluorescence in situ hybridization (FISH). Fungal DNA was found in 35 of 38 (92.11%) coronary heart disease patients by either polymerase chain reaction (PCR) with universal primers or in situ hybridization analysis (n = 5), but not in any control sample. In a clone library with more than 350 sequenced clones from pooled patient DNA, an overall richness of 19 different fungal phylotypes could be observed. Fungal profiles of coronary heart disease patients obtained by DGGE analysis showed a median richness of fungal species of 5 (range from 2 to 9) with a high interindividual variability (mean similarity 18.83%). For the first time, the presence of fungal components in atherosclerotic plaques has been demonstrated. Coronary atheromatous plaques harbour diverse and variable fungal communities suggesting a polymicrobial contribution to the chronic inflammatory aetiology.
机译:在冠状动脉斑块中发现了细菌DNA,因此可以得出结论,细菌可能是冠状动脉粥样硬化背后的慢性炎症过程的触发因素。然而,微生物谱是复杂的,除了细菌以外的微生物是否参与冠心病尚不清楚。使用克隆文库,变性梯度凝胶分析(DGGE),原位杂交和荧光原位杂交(FISH)在38例患者和对照(未受影响的冠状动脉)的基于导管的斑块切除术中获得的动脉粥样硬化组织中系统地研究了真菌18S rDNA签名。通过通用引物的聚合酶链反应(PCR)或原位杂交分析(n = 5)在38名冠心病患者中的35名(92.11%)中发现了真菌DNA,但在任何对照样品中均未发现。在具有来自合并的患者DNA的350多个测序克隆的克隆文库中,可以观察到19种不同真菌系统型的总体丰富度。通过DGGE分析获得的冠心病患者的真菌谱显示,真菌种类的中位数丰富度为5(从2到9),且个体间差异很大(平均相似度为18.83%)。首次证明了动脉粥样硬化斑块中存在真菌成分。冠状动脉粥样斑块具有多种多样的真菌群落,提示对慢性炎症病因有多种微生物作用。

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