Abstract
Invasive aspergillosis (IA) is an important cause of morbidity and mortality among immunocompromised people. Imaging and specimen tests used in the clinical diagnosis of aspergillosis with weak and indistinct defects leads to delay in the treatment of early aspergillosis patients. The developing molecular techniques provide a new method for the aspergillosis diagnosis. However, the existing methods are complex, time-consuming and may even be potentially hazardous. In this study, we developed a simple and rapid Aspergillus fumigatus spores DNA isolation assay using synthesized zinc oxide (ZnO). ZnO nanoparticles were used to take the place of the traditional commercial lysis buffer. The quality and quantity of the extracted DNA were sufficient for further diagnostics with polymerase chain reaction (PCR) analysis. This method offers easy, green, and economic alternative DNA isolation for the diagnosis of invasive aspergillosis.
Highlights
Invasive aspergillosis (IA) plays a significant role in the morbidity and mortality of immunocompromised individuals, bone marrow transplant recipients, cancer patients, human immuno-deficiency virus (HIV) patients, and patients undergoing treatment with immunomodulators [1,2]
A. fumigatus is common in all environments but is difficult to distinguish from certain other molds (A. flavus, A. niger, and A. terreus) under the microscope [3,4]
The most common tests used to diagnose aspergillosis are the imaging tests using a chest X-ray or computerized tomography (CT) and specimen tests of sputum, tissue, and blood [5,6,7]
Summary
Invasive aspergillosis (IA) plays a significant role in the morbidity and mortality of immunocompromised individuals, bone marrow transplant recipients, cancer patients, human immuno-deficiency virus (HIV) patients, and patients undergoing treatment with immunomodulators [1,2]. Most commercial fungal DNA extraction kits used in clinics and laboratories are based on a standard method comprising the lyophilization of mycelia, disruption of the cell wall by grinding, extraction of DNA in a buffer containing sodium dodecyl sulfate, removal of proteins with a mixture of phenol and chloroform, and precipitation of DNA with 2-propanol [12,13,14]. We first report an assay for fungal DNA isolation based on the effectiveness and performance of a cell lysis buffer at room temperature without additional instruments. The performance of this ZnO-based fungal DNA isolation assay is superior to those of other two kinds of commercial ZnO types, ZnO-C-100 and ZnO-C-5000
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