Nucleic acid sequence-based amplification of Aspergillus RNA in blood samples.
Nucleic acid sequence-based amplification (NASBA), an isothermal amplification technique, was established and evaluated for the detection of Aspergillus RNA and compared with a previously published, well-defined real-time PCR assay amplifying a region of the Aspergillus 18S rRNA gene. NASBA showed a lower detection limit of 1 CFU and detected RNA from five different clinically relevant Aspergillus species, including Aspergillus fumigatus. All 77 blood samples tested by PCR and NASBA showed identical results in both assays. Results with the NASBA technique were obtained within 6 h. Thus, the NASBA technique provided a valuable tool for sensitive, specific, fast, and reliable detection of Aspergillus RNA with potential for routine diagnosis, including the possibility to test the viability of cells.
Medizinische Klinik, Abteilung II, Eberhard-Karls-Universität Tübingen, 72076 Tübingen, Germany. juergen.loeffler@med.uni-tuebingen.de
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Loeffler
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Hebart
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Cox
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Flues
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Schumacher
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Einsele
D001228:Aspergillosis / Q000382:microbiology*; D001230:Aspergillus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D015169:Colony Count, Microbial; D016469:Fungemia / Q000382:microbiology; D006801:Humans; D016133:Polymerase Chain Reaction; D012331:RNA, Fungal / Q000097:blood*; D021161:Self-Sustained Sequence Replication; D012680:Sensitivity and Specificity
D012331:RNA, Fungal