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Comparison of genotyping methods for Cunninghamella bertholletiae Cunninghamella bertholletiae

机译:Cunninghamella Bertholletiae Cunninghamella Bertholletia的基因分型方法比较

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Summary Background Invasive fungal infections caused by filamentous fungi of the order Mucorales are serious complications in immunocompromised patients and often associated with fatal outcome. As a member of this order, Cunninghamella bertholletiae is a saprophytic fungus with naturally exhibited high minimum inhibitory concentrations against common antifungal drugs and with the potential for outbreaks in clinical settings. Objectives and methods In a proof‐of‐principle study, we evaluated the performance of microsatellite markers for the discrimination of thirteen C.?bertholletiae isolates from various sources in comparison with a repetitive sequence‐based PCR (rep‐ PCR ) and random amplification of polymorphic DNA ( RAPD ). Based on the higher discriminatory power of the microsatellite PCR with five separate primer pairs (Simpson's index of 1 vs 0 [ RAPD ] and 0 [rep‐ PCR ]), the novel method was applied to eight additional isolates, including four well‐characterised isolates from a cluster of infections in a next step. Results In total, microsatellite PCR identified 21 separate genotypes. A probable epidemiological association of the cluster isolates could be demonstrated by microsatellite genotyping. Conclusion In conclusion, our findings demonstrate the value of microsatellite PCR in genotyping Cunninghamella bertholletiae and its potential for future applications with other species of the order Mucorales.
机译:发明内容背景下霉菌霉菌引起的侵袭性真菌感染是免疫抑制患者的严重并发症,通常与致命结果相关。作为本次订单的成员,Cunninghamella Bertholletiae是一种嗜酸性真菌,其天然存在对常见抗真菌药物的最低最低抑制浓度,以及临床环境中爆发的可能性。在原则上的验证中的目标和方法,我们评估了微卫星标志物的性能与各种来源的十三C.?bertholletiae分离株的判别的性能与基于重复的序列的PCR(Rep-PCR)和随机扩增相比多态性DNA(RAPD)。基于微卫星PCR的较高辨别力,用五个单独的引物对(SIMPSON的1 Vs 0 [RAPD]和0 [REP-PCR]),将新方法应用于八个另外的分离株,包括四个特征性的分离物从下一步中的一群感染。结果总,微卫星PCR鉴定了21种单独的基因型。通过微卫星基因分型可以证明簇分离物的可能的流行病学结合。结论得出结论,我们的研究结果证明了微卫星PCR在基因分型康宁米拉Bertholletia的价值及其未来应用的潜力与其他物种粘液粘膜。

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