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首页> 外文期刊>Journal of Clinical Microbiology >Multilocus DNA Sequence Comparisons Rapidly Identify Pathogenic Molds
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Multilocus DNA Sequence Comparisons Rapidly Identify Pathogenic Molds

机译：多基因座DNA序列比较可快速鉴定致病菌

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The increasing incidence of opportunistic fungal infections necessitates rapid and accurate identification of the associated fungi to facilitate optimal patient treatment. Traditional phenotype-based identification methods utilized in clinical laboratories rely on the production and recognition of reproductive structures, making identification difficult or impossible when these structures are not observed. We hypothesized that DNA sequence analysis of multiple loci is useful for rapidly identifying medically important molds. Our study included the analysis of the D1/D2 hypervariable region of the 28S ribosomal gene and the internal transcribed spacer (ITS) regions 1 and 2 of the rRNA operon. Two hundred one strains, including 143 clinical isolates and 58 reference and type strains, representing 43 recognized species and one possible new species, were examined. We generated a phenotypically validated database of 118 diagnostic alleles. DNA length polymorphisms detected among ITS1 and ITS2 PCR products can differentiate 20 of 33 species of molds tested, and ITS DNA sequence analysis permits identification of all species tested. For 42 of 44 species tested, conspecific strains displayed >99% sequence identity at ITS1 and ITS2; sequevars were detected in two species. For all 44 species, identifications by genotypic and traditional phenotypic methods were 100% concordant. Because dendrograms based on ITS sequence analysis are similar in topology to 28S-based trees, we conclude that ITS sequences provide phylogenetically valid information and can be utilized to identify clinically important molds. Additionally, this phenotypically validated database of ITS sequences will be useful for identifying new species of pathogenic molds.
机译：机会性真菌感染的发病率不断增加，因此需要快速准确地鉴定相关真菌，以促进患者的最佳治疗。临床实验室中使用的传统的基于表型的鉴定方法依赖于生殖结构的产生和识别，如果未观察到这些结构，则很难或不可能进行鉴定。我们假设多个基因座的DNA序列分析可用于快速识别具有医学重要性的霉菌。我们的研究包括对28S核糖体基因的D1 / D2高变区以及rRNA操纵子的内部转录间隔区（ITS）1和2的分析。检验了代表43种公认物种和一种可能的新物种的201个菌株，包括143个临床分离株和58个参考和类型菌株。我们生成了118个诊断等位基因的表型验证数据库。在ITS1和ITS2 PCR产物中检测到的DNA长度多态性可以区分33种受测霉菌中的20种，而ITS DNA序列分析可以鉴定所有受测霉菌。对于测试的44个物种中的42个，同种菌株在ITS1和ITS2上显示出> 99％的序列同一性。在两个物种中发现了sequevars。对于所有44个物种，通过基因型和传统表型方法进行的鉴定都是100％一致的。因为基于ITS序列分析的树状图在拓扑结构上与基于28S的树相似，所以我们得出结论，ITS序列提供了系统上有效的信息，可用于识别具有临床意义的霉菌。另外，这个表型验证的ITS序列数据库将对识别新的致病性霉菌有用。

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