Evaluation of nucleic acid sequence based amplification using fluorescence resonance energy transfer (FRET-NASBA) in quantitative detection of Aspergillus 18S rRNA

被引：10

作者：

Park, Chulmin ^{[2
]}

Kwon, Eun-Young ^{[2
]}

Shin, Na-Young ^{[2
]}

Choi, Su-Mi ^{[1
]}

Kim, Si-Hyun ^{[1
]}

Park, Sun Hee ^{[1
]}

Lee, Dong-Gun ^{[1
]}

Choi, Jung-Hyun ^{[1
]}

Yoo, Jin-Hong ^{[1
]}

机构：

[1] Catholic Univ Korea, Coll Med, Dept Internal Med, Seoul, South Korea

[2] Catholic Univ Korea, Coll Med, Catholic Res Inst Med Sci, Seoul, South Korea

来源：

MEDICAL MYCOLOGY | 2011年 / 49卷 / 01期

关键词：

Aspergillus; NASBA; FRET; REAL-TIME DETECTION; INVASIVE ASPERGILLOSIS; DIAGNOSIS; QUANTIFICATION; ASSAY;

D O I：

10.3109/13693786.2010.507604

中图分类号：

R51 [传染病];

学科分类号：

100401 ;

摘要：

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<SU-2</SU amol) than the FRET. FRET-NASBA showed a sensitivity of 10<SU-1</SU 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.</.

引用

页码：73 / 79

页数：7

共 21 条

[1] Detection and quantification of the red tide dinoflagellate Karenia brevis by real-time nucleic acid sequence-based amplification [J].