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首页> 外文期刊>Journal of Clinical Microbiology >Automated Extraction of Genomic DNA from Medically Important Yeast Species and Filamentous Fungi by Using the MagNA Pure LC System
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Automated Extraction of Genomic DNA from Medically Important Yeast Species and Filamentous Fungi by Using the MagNA Pure LC System

机译：通过使用MagNA纯LC系统自动从医学上重要的酵母物种和丝状真菌中提取基因组DNA

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A fully automated assay was established for the extraction of DNA from clinically important fungi by using the MagNA Pure LC instrument. The test was evaluated by DNA isolation from 23 species of yeast and filamentous fungi and by extractions (n = 28) of serially diluted Aspergillus fumigatus conidia (105 to 0 CFU/ml). Additionally, DNA from 67 clinical specimens was extracted and compared to the manual protocol. The detection limit of the MagNA Pure LC assay of 10 CFU corresponded to the sensitivity when DNA was extracted manually; in 9 of 28 runs, we could achieve a higher sensitivity of 1 CFU/ml blood, which was found to be significant (p ≤ 0.004). DNA from all fungal species analyzed could be extracted and amplified by real-time PCR. Negative controls from all MagNA Pure isolations remained negative. Sixty-three clinical samples showed identical results by both methods, whereas in 4 of 67 samples, discordant results were obtained. Thus, the MagNA Pure LC technique offers a fast protocol for automated DNA isolation from numerous fungi, revealing high sensitivity and purity.
机译：通过使用MagNA Pure LC仪器，建立了用于从临床重要真菌中提取DNA的全自动测定方法。通过从23种酵母和丝状真菌中进行DNA分离以及连续稀释的烟曲霉（ems）的分生孢子（10 5 < / sup>到0 CFU / ml）。此外，从67个临床标本中提取DNA并与手册进行比较。 MagNA Pure LC法检测10 CFU的检出限与手动提取DNA时的灵敏度相对应。在28次运行中的9次中，我们可以达到1 CFU / ml血液的更高灵敏度，这被认为是显着的（p≤0.004）。可以通过实时PCR提取和扩增来自所有真菌物种的DNA。来自所有MagNA Pure分离株的阴性对照仍为阴性。两种方法的63个临床样品显示出相同的结果，而67个样品中有4个获得不一致的结果。因此，MagNA Pure LC技术提供了一种从多种真菌中自动分离DNA的快速方案，具有很高的灵敏度和纯度。

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