title: Evaluation of nucleic acid sequence based amplification using fluorescence resonance energy transfer (FRET-NASBA) in quantitative detection of Aspergillus 18S rRNA.
abstract: 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.
authors: Catholic Research Institutes of Medical Science, The Catholic University of Korea, College of Medicine, Seoul, South Korea.
authors: Chulmin
authors: C
authors: Park
authors: Eun-Young
authors: EY
authors: Kwon
authors: Na-Young
authors: NY
authors: Shin
authors: Su-Mi
authors: SM
authors: Choi
authors: Si-Hyun
authors: SH
authors: Kim
authors: Sun Hee
authors: SH
authors: Park
authors: Dong-Gun
authors: DG
authors: Lee
authors: Jung-Hyun
authors: JH
authors: Choi
authors: Jin-Hong
authors: JH
authors: Yoo
mesh_terms: D001228:Aspergillosis / Q000175:diagnosis*; D001230:Aspergillus / Q000235:genetics / Q000302:isolation & purification*; D017931:DNA Primers / Q000235:genetics; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D006801:Humans; D009172:Mycology / Q000379:methods*; D015345:Oligonucleotide Probes / Q000235:genetics; D012331:RNA, Fungal / Q000235:genetics*; D012337:RNA, Ribosomal, 18S / Q000235:genetics*; D021161:Self-Sustained Sequence Replication / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D017931:DNA Primers; D015345:Oligonucleotide Probes; D012331:RNA, Fungal; D012337:RNA, Ribosomal, 18S