Assessment of Aspergillus fumigatus in Guinea Pig Bronchoalveolar Lavages and Pulmonary Tissue by Culture and Realtime Polymerase Chain Reaction Studies
| Autor: | Nathan P. Wiederhold, Laura K. Najvar, Thomas F. Patterson, Frederick T. Guilford, David C. Straus, William R. Kirkpatrick, John S. Sutton, Dennis Hooper, Vincent E. Bolton |
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| Jazyk: | angličtina |
| Rok vydání: | 2012 |
| Předmět: |
Male
Guinea Pigs bronchoalveolar lavages Real-Time Polymerase Chain Reaction Aspergillosis Article Catalysis law.invention Microbiology Aspergillus fumigatus Realtime PCR lcsh:Chemistry Inorganic Chemistry Guinea pig chemistry.chemical_compound law medicine Animals Physical and Theoretical Chemistry DNA Fungal Lung lcsh:QH301-705.5 Molecular Biology Spectroscopy Polymerase chain reaction DNA Primers Detection limit biology Organic Chemistry General Medicine biology.organism_classification medicine.disease Molecular biology Computer Science Applications Real-time polymerase chain reaction lcsh:Biology (General) lcsh:QD1-999 chemistry Primer (molecular biology) Bronchoalveolar Lavage Fluid DNA |
| Zdroj: | International Journal of Molecular Sciences Volume 13 Issue 1 Pages 726-736 International Journal of Molecular Sciences, Vol 13, Iss 1, Pp 726-736 (2012) |
| ISSN: | 1422-0067 |
| DOI: | 10.3390/ijms13010726 |
| Popis: | 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 detection limit of the assay ranged from 6 × 10(4) copies to 6 copies. Since blood cultures are rarely positive for Aspergillosis, our data indicate that qualitative Realtime PCR, in combination with the appropriate DNA primers and probe can serve as an effective diagnostic tool in the early detection of fungal infections. |
| Databáze: | OpenAIRE |
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