Rapid characterisation and detection of Aspergillus fumigatus and Aspergillus flavus from environmental air samples

• Main Author: Chiew, Jean Ling
• Other Authors: Yap Peng Huat Eric
• Format: Final Year Project
• Language: English
• Published: 2017
• Subjects: DRNTU::Science::Medicine
• Online Access: http://hdl.handle.net/10356/72625
• Institution: Nanyang Technological University

Aspergillus causes life-threatening infections in certain at-risk patient populations, and is linked to millions of asthma or other allergic syndromes each year. Though there is fair understanding of Aspergillus detection from clinical specimens, little has been published on detection of airborne Aspergillus to assess risk of exposure from spores in Singapore’s urban indoor environment. Furthermore, while Polymerase Chain Reaction techniques have proven high sensitivity and specificity over culture-based methods for detecting Aspergillus, they lack global standardisation and hence are not routinely used clinically. When used, traditional methods such as conventional or Real-Time PCR (RTPCR) take up to hours to detect presence of Aspergillus, potentially delaying diagnosis or treatment. As such, this paper’s objective is “rapid characterisation and detection of Aspergillus fumigatus and Aspergillus flavus from environmental samples”. This paper has succeeded in using conventional PCR and RT-PCR to detect and identify Aspergillus fumigatus and Aspergillus flavus from environmental air samples of significance to Singaporean students, working and laboratory personnel. Samples were identified successfully both at genus and species level, analysed using gel electrophoresis and melt curves. We have also successfully developed Fast PCR equipment and techniques to amplify Aspergillus in under 10 minutes. Detection is complemented by gel electrophoresis and fluorescence comparisons preand post-Fast PCR. With further development, this new technology can prospectively be used for rapid Aspergillus detection in clinical specimens for point-of-care diagnostics, aiding diagnosis and treatment in a bid to reduce Aspergillus-related mortality and morbidity.