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首页> 美国卫生研究院文献>Journal of Clinical Microbiology >New Polymorphic Microsatellite Markers Able To Distinguish among Candida parapsilosis Sensu Stricto Isolates
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New Polymorphic Microsatellite Markers Able To Distinguish among Candida parapsilosis Sensu Stricto Isolates

机译：新的多态微卫星标记能够区分假丝酵母念珠菌分离株

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Among the Candida species causing bloodstream infections, Candida parapsilosis is one of the most frequently isolated. The objective of the present work was the identification of new microsatellite loci able to distinguish among C. parapsilosis isolates. DNA sequences with trinucleotide repeats were selected from the C. parapsilosis genome database. PCR primer sets flanking the microsatellite repeats were designed and tested with 20 independent isolates. On the basis of the amplification efficiency, specificity, and observed polymorphism, four of the sequences were selected for strain typing. Two hundred thirty-three independent C. parapsilosis sensu stricto isolates were genotyped by using these markers. The polymorphic loci exhibited from 20 to 42 alleles and 39 to 92 genotypes. In a multiplex analysis, 192 genotypes were obtained and the combined discriminatory power of the four microsatellites was 0.99. Reproducibility was demonstrated by submission of subcultures of 4 isolates each, in triplicate, interspersed with unique numbers among a group of 30 isolates for blind testing. Comparison of the genotypes obtained by microsatellite analysis and those obtained by randomly amplified polymorphic DNA analysis, restriction fragment length polymorphism analysis, and internal transcribed sequence grouping was performed and showed that the microsatellite method could distinguish individual isolates; none of the other methods could do that. Related species, C. orthopsilosis and C. metapsilosis, were not confused with C. parapsilosis sensu stricto. These new microsatellites are a valuable tool for use for the differentiation of C. parapsilosis sensu stricto strains, vital in epidemiology to answer questions of strain relatedness and determine pathways of transmission.
机译：在引起血液感染的念珠菌中，副念珠菌是最常见的一种。本工作的目的是鉴定新的微卫星基因座，该基因座能够区分副枝梭菌。具有三核苷酸重复序列的DNA序列选自副枝隐孢子虫基因组数据库。设计了微卫星重复序列两侧的PCR引物，并用20个独立的分离物进行了测试。基于扩增效率，特异性和观察到的多态性，选择了四个序列用于菌株分型。使用这些标记对233个独立的C. parapsilosis sensu stricto分离株进行了基因分型。多态性位点表现出20至42个等位基因和39至92个基因型。在多重分析中，获得了192个基因型，四个微卫星的组合判别力为0.99。通过提交4个分离株的亚培养物，一式三份，并散布在一组30个分离株中进行盲测的唯一编号，证明了可重复性。比较了通过微卫星分析获得的基因型与通过随机扩增多态性DNA分析，限制性片段长度多态性分析和内部转录序列分组获得的基因型，结果表明，微卫星方法可以区分单个菌株。其他方法都无法做到这一点。相关物种，C。orthopsilosis和C. metapsilosis，没有与C. parapsilosis sensu stricto混淆。这些新的微卫星是用于区分短小隐孢子虫严密菌株的有价值的工具，在流行病学中至关重要，以回答菌株相关性的问题并确定传播途径。

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