Catholic Research Institutes of Medical Science, The Catholic University of Korea, College of Medicine, Seoul, South Korea.
Med Mycol. 2011 Jan;49(1):73-9. doi: 10.3109/13693786.2010.507604. Epub 2010 Aug 18.
We attempted to apply fluorescence resonance energy transfer technology to nucleic acid sequence-based amplification (FRET-NASBA) on the platform of the LightCycler system to detect Aspergillus species. Primers and probes for the Aspergillus 18S rRNA were newly designed to avoid overlapping with homologous sequences of human 18s rRNA. NASBA using molecular beacon (MB) showed non-specific results which have been frequently observed from controls, although it showed higher sensitivity (10(-2) amol) than the FRET. FRET-NASBA showed a sensitivity of 10(-1) amol and a high fidelity of reproducibility from controls. As FRET technology was successfully applied to the NASBA assay, it could contribute to diverse development of the NASBA assay. These results suggest that FRET-NASBA could replace previous NASBA techniques in the detection of Aspergillus.
我们试图将荧光共振能量转移技术（FRET）应用于核酸序列扩增（NASBA）平台，以检测曲霉属物种。为了避免与人类 18s rRNA 的同源序列重叠，我们新设计了用于曲霉 18S rRNA 的引物和探针。使用分子信标（MB）的 NASBA 显示出非特异性结果，尽管它的灵敏度（10(-2) amol）比 FRET 高。FRET-NASBA 的灵敏度为 10(-1) amol，从对照中显示出很高的重现性和准确性。由于 FRET 技术已成功应用于 NASBA 检测，因此它可以为 NASBA 检测的多样化发展做出贡献。这些结果表明，FRET-NASBA 可以替代以前的 NASBA 技术来检测曲霉属。
