Use of a novel panel of nine short tandem repeats for exact and high-resolution fingerprinting of Aspergillus fumigatus isolates
Use of a novel panel of nine short tandem repeats for exact and high-resolution fingerprinting of Aspergillus fumigatus isolates
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DOI：
10.1128/jcm.43.8.4112-4120.2005
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发表时间：
2005-08-01
影响因子：
9.4
通讯作者：
Klaassen, CHW
中科院分区：
文献类型：
--
作者：
de Valk, HA;Meis, JFGM;Klaassen, CHW
在这里，我们描述了一个新的面板的短串联重复序列（STR）的一种新的精确分型检测，可用于区分烟曲霉菌株。从现有的基因组A中筛选出9个STR标记。fumigatus序列，并分为三个重复的多重PCR。每个多重反应分别扩增三个二、三或四核苷酸重复。所有9个STIR标记用于分析100个可能无关的A。烟曲霉菌株对于每个标记，在该群体中发现11至37个等位基因。一个分离株被证明是至少两种不同分离株的混合物。与其余99个分离株，获得96个不同的指纹图谱。单个标记的Simpson多样性指数在0.77 ~ 0.97之间。二，三，和四核苷酸重复的多重组合的多样性指数范围从0.9784到0.9968。所有9个标记的组合产生了辛普森的多样性指数为0.9994，表明这些新的位点的高区分能力。理论上，这组标记能够区分不少于27 × 10（9）种不同的基因型。多重PCR方法允许在短时间内测试大量标记。该检测试剂盒的精确性质结合了高重现性和易于交换的结果，使其成为大规模流行病学研究的非常合适的工具。
Here we describe a new panel of short tandem repeats (STRs) for a novel exact typing assay that can be used to discriminate between Aspergillus fumigatus isolates. A total of nine STR markers were selected from available genomic A. fumigatus sequences and were divided into three multicolor multiplex PCRs. Each multiplex reaction amplified three di-, tri-, or tetranucleotide repeats, respectively. All nine STIR markers were used to analyze 100 presumably unrelated A. fumigatus isolates. For each marker, between 11 and 37 alleles were found in this population. One isolate proved to be a mixture of at least two different isolates. With the remaining 99 isolates, 96 different fingerprinting profiles were obtained. The Simpson's diversity index for the individual markers ranged from 0.77 to 0.97. The diversity index for the multiplex combination of di-, tri-, and tetranucleotide repeats ranged from 0.9784 to 0.9968. The combination of all nine markers yielded a Simpson's diversity index of 0.9994, indicative of the high discriminatory power of these new loci. In theory, this panel of markers is able to discriminate between no less than 27 x 10(9) different genotypes. The multicolor multiplex approach allows large numbers of markers to be tested in a short period of time. The exact nature of the assay combines high reproducibility with the easy exchange of results and makes it a very suitable tool for large-scale epidemiological studies.