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首页> 外文期刊>Japanese journal of infectious diseases >Comparison of whole genome amplification methods for detecting pathogenic bacterial genomic DNA using microarray.
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Comparison of whole genome amplification methods for detecting pathogenic bacterial genomic DNA using microarray.

机译：使用微阵列检测病原细菌基因组DNA的全基因组扩增方法的比较。

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摘要

The genetic diagnosis of pathogenic agents using microarrays has the advantage of high-throughput detection, but a relatively large amount of DNA sample is required. To obtain a sufficient amount of DNA for molecular diagnoses, several whole genome amplification (WGA) methods have been proposed. In this study, using Francisella tularensis and Escherichia coli as models, we compared four WGA methods in terms of their efficiency of amplification of whole genomic DNA in order to identify the most suitable method for preparing DNA to be used for microarray analysis. It was possible to obtain more than 1.5 microg of products from 10 ng of F. tularensis and E. coli genomic DNA using four methods, but biases in the amplification of bacterial genes were least prominent in the multiple displacement amplification (MDA) or OmniPlex WGA. When the amplified DNAs were applied to microarray slides consisting of 32 different genes probes, DNAs amplified by Phi29 v2 of MDA and OmniPlex WGA showed high signal intensity as well as a high signal-to-noise ratio for all 32 genes. These results indicate that Phi29 v2 and OmniPlex WGA are useful methods for obtaining sufficient DNA from a limited amount of samples for the detection of microbes using microarrays.
机译：使用微阵列对病原体进行遗传诊断具有高通量检测的优势，但是需要相对大量的DNA样品。为了获得足够的DNA用于分子诊断，已经提出了几种全基因组扩增（WGA）方法。在这项研究中，使用土拉弗朗西斯菌和大肠杆菌作为模型，我们比较了四种WGA方法在扩增整个基因组DNA方面的效率，从而确定了最合适的制备用于微阵列分析的DNA的方法。使用四种方法可以从10 ng的F. tularensis和大肠杆菌基因组DNA中获得超过1.5 microg的产物，但是在多重置换扩增（MDA）或OmniPlex WGA中，细菌基因扩增的偏差最少。 。当将扩增的DNA应用于由32种不同基因探针组成的微阵列玻片时，通过MDA的Phi29 v2和OmniPlex WGA扩增的DNA对所有32个基因均显示出高信号强度和高信噪比。这些结果表明，Phi29 v2和OmniPlex WGA是从有限数量的样品中获得足够的DNA以便使用微阵列检测微生物的有用方法。

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