Int. J. Mol. Sci. 2012, 13, 726-736; doi:10.3390/ijms13010726 International Journal of Molecular Sciences ISSN 1422-0067 www.mdpi.com/journal/ijms Article Assessment of Aspergillus fumigatus in Guinea Pig Bronchoalveolar Lavages and Pulmonary Tissue by Culture and Realtime Polymerase Chain Reaction Studies Dennis G. Hooper 1, *, Vincent E. Bolton 1 , John S. Sutton 2 , Frederick T. Guilford 3 , David C. Straus 4 , Laura K. Najvar 5,6 , Nathan P. Wiederhold 5,7 , William R. Kirkpatrick 5,6 and Thomas F. Patterson 5,6 1 RealTime Laboratories, Inc, 4100 Fairway Court, #600, Carrollton, TX 75010, USA 2 S&S BioConsulting, LLC, Austin, TX 78660, USA 3 Your Energy Systems, 5050 El Camino Real, #110, Los Altos, CA 94022, USA 4 Department of Microbiology and Immunology, Texas Tech University Health Sciences Center, Lubbock, TX 79430, USA 5 Department of Medicine, The University of Texas Health Science Center at San Antonio, San Antonio, TX 78229, USA 6 South Texas Veterans Health Care System, San Antonio, TX 78229, USA 7 Division of Pharmacotherapy, University of Texas at Austin College of Pharmacy, Austin, TX 78712, USA * Author to whom correspondence should be addressed; E-Mail: dhooper@realtimelab.com; Tel.: +1-972-243-7754; Fax: +1-972-243-7759. Received: 24 November 2011; in revised form: 10 December 2011 / Accepted: 6 January 2012 / Published: 11 January 2012 Abstract: In this study we pursued a diagnostic target in Aspergillus fumigatus (AF) by using qualitative Realtime PCR combined with proprietary DNA primers and a hydrolysis probe specific for this fungal target. Qualitative Realtime PCR is a diagnostic tool that utilizes Realtime PCR technology and detects the presence or absence target specific DNA within a predetermined detection range. Respiratory tissue and fluids from experimentally infected guinea pigs were tested by extracting DNA from the samples which were amplified and detected using AF specific DNA primers and probe. This study included qualitative evaluations of all specimens for the presence of the DNA of AF. The findings in the tissues after AF infection were compared to the numbers of spores in aerosolized samples used to inoculate the animals. Results demonstrated that the specific probe and primer set could detect the presence or absence of AF DNA in the sample. The qualitative OPEN ACCESS