Development of Nested-PCR Assay for The Rapid Detection of Aspergillus fumigatus

Aspergillus fumigatus is a fungal pathogen that causes Aspergillosis. This infection can develop rapidly in the human body and often causes death for immunocompromised people. Aspergillus's conidia are easily distributed in the environment and enter the paranasal sinuses or lungs. Identification of A. fumigatus is very important as an effort to prevent the spread of this fungal pathogen early. A molecular-based rapid method is promised to identify A. fumigatus without relying on morphological observations. This study represents the initial process to achieve a consensus method of nested Polymerase Chain Reaction (PCR) to diagnose A. fumigatus infection. The methods were implemented include the collection of A. fumigatus isolates, macroscopic and microscopic characterization. DNA isolation were carried out to obtain the total genome of A. fumigatus. Primers were designed to amplify 630 bp of internal transcribed spacer (ITS). To enhance the sensitivity, another primer pair was designed to amplify a 400 bp region of ITS gene and was used in a nested PCR assay. In conclusion, this initial process of nested PCR may be utilized to detect fungal A. fumigatus which can aid the diagnosis of invasive fungal disease.