Detection of Aspergillus fumigatus in a rat model of invasive pulmonary aspergillosis by real-time nucleic acid sequence-based amplification.

Journal of Clinical Microbiology
Yanan ZhaoDavid S Perlin

Abstract

Rapid and sensitive detection of Aspergillus from clinical samples may facilitate the early diagnosis of invasive pulmonary aspergillosis (IPA). A real-time nucleic acid sequence-based amplification (NASBA) method was investigated by use of an inhalational rat model of IPA. Immunosuppressed male Sprague-Dawley rats were exposed to Aspergillus fumigatus spores for an hour in an aerosol chamber. Bronchoalveolar lavage (BAL) fluid, lung tissues, and whole blood were collected from five infected rats at 1, 24, 48, 72, and 96 h postinfection and five uninfected rats at the end of the experiment. Total nucleic acid (TNA) was extracted on an easyMAG instrument. A primer-molecular beacon set targeting 28S rRNA was designed to detect Aspergillus spp. The results were compared to those of quantitative PCR (qPCR) (18S rDNA) and quantitative culture. The analytical sensitivity of the real-time NASBA assay was <1 CFU/assay. A linear range of detection was demonstrated over 5 log units of conidia (10 to 10(5) spores). Both NASBA and qPCR showed a progressive increase in lung tissue burdens, while the CFU counts were stable over time. The fungal burdens in BAL fluid were more variable and not indicative of a progressive infection. The results...Continue Reading

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Citations

Jul 3, 2013·Immunobiology·Amal Atia Mhfuod El-MuzghiMilena Kataranovski
Mar 7, 2013·The Journal of Antimicrobial Chemotherapy·Yanan ZhaoMaiken C Arendrup
Jul 19, 2012·PloS One·Gemma L JohnsonStephen A Bustin
Aug 13, 2010·Clinical Medicine Insights. Oncology·Rodrigo Lopes da SilvaAida Botelho de Sousa
Nov 26, 2013·Future Microbiology·Sean X Zhang
Oct 17, 2012·Clinics in Dermatology·Jan SpringerJuergen Loeffler
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Jul 3, 2013·American Journal of Rhinology & Allergy·Fang ZhangChangqing Zhao
Aug 18, 2018·Veterinary Clinical Pathology·Masha G SavelieffPanagiotis Azmanis
Jun 1, 2017·Frontiers in Microbiology·Guillaume Desoubeaux, Carolyn Cray
Oct 2, 2019·Journal of Fungi·Amir ArastehfarTeun Boekhout
Jun 24, 2016·Microbiology Spectrum·Margaret V Powers-FletcherKimberly E Hanson

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