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用于检测曲霉菌的分子诊断平台。

Molecular diagnostic platforms for detecting Aspergillus.

作者信息

Perlin David S, Zhao Yanan

机构信息

Public Health Research Institute, Department of Microbiology and Molecular Genetics, UMDNJ-New Jersey Medical School, Newark, New Jersey 07103, USA.

出版信息

Med Mycol. 2009;47 Suppl 1:S223-32. doi: 10.1080/13693780802126583. Epub 2008 Jun 6.

DOI:10.1080/13693780802126583
PMID:18608902
Abstract

Invasive aspergillosis remains difficult to diagnose despite advances in imaging and antigen-based serological testing. To overcome this problem, nucleic acid (NA)-based amplification assays were introduced to identify infecting pathogens. Unfortunately, the reliability of such assays to detect Aspergillus spp. has met with mixed success. A new generation of NA platforms are emerging, which greatly improve our ability to detect Aspergillus-specific DNA and RNA from respiratory and blood samples. These platforms can accurately detect a single genome, and the emergence of pan-fungal and pan-Aspergillus probes offer promise for broader detection. PCR remains the most important platform, especially when coupled with real-time probes. It is multiplex friendly and can distinguish between closely related target sequences. Nucleic acid sequence-based amplification (NASBA) is an RNA-directed isothermal transcription-based amplification platform, which is more robust than PCR resulting in a 10(14)-fold amplification. RNA-based detection facilitates more target options and can be used to assess cell viability. Both DNA and RNA amplification platforms take advantage of allele-specific properties of probes, which are valuable for assessing drug resistance markers. Finally, as new molecular diagnostic platforms mature, their role may expand to include early monitoring of therapy.

摘要

尽管在影像学和基于抗原的血清学检测方面取得了进展,但侵袭性曲霉病仍然难以诊断。为克服这一问题,引入了基于核酸(NA)的扩增检测方法来鉴定感染病原体。不幸的是,此类检测方法检测曲霉属菌种的可靠性参差不齐。新一代的NA平台正在出现,这极大地提高了我们从呼吸道和血液样本中检测曲霉特异性DNA和RNA的能力。这些平台能够准确检测单个基因组,泛真菌和泛曲霉探针的出现为更广泛的检测带来了希望。聚合酶链反应(PCR)仍然是最重要的平台,尤其是与实时探针结合使用时。它有利于多重检测,并且能够区分密切相关的靶序列。基于核酸序列的扩增(NASBA)是一种基于RNA定向等温转录的扩增平台,比PCR更强大,可实现10¹⁴倍的扩增。基于RNA的检测提供了更多的靶标选择,可用于评估细胞活力。DNA和RNA扩增平台都利用了探针的等位基因特异性特性,这对于评估耐药标记物很有价值。最后,随着新的分子诊断平台逐渐成熟,它们的作用可能会扩大到包括治疗的早期监测。

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Molecular diagnostic platforms for detecting Aspergillus.用于检测曲霉菌的分子诊断平台。
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