Perlin David S, Zhao Yanan
Public Health Research Institute, Department of Microbiology and Molecular Genetics, UMDNJ-New Jersey Medical School, Newark, New Jersey 07103, USA.
Med Mycol. 2009;47 Suppl 1:S223-32. doi: 10.1080/13693780802126583. Epub 2008 Jun 6.
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.
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。