Detection of Azole Drug Resistance of the Aspergillus Species Cyp51a Gene by PCR

• Main Authors: Raksha, Gurjeet Singh, A D Urhekar
• Format: Article
• Language: English
• Published: ADICHUNCHANAGIRI INSTITUTE OF MEDICAL SCIENCES 2019-06-01
• Series: Journal of Medical Sciences and Health
• Subjects: azole; polymerase chain reaction; gene sequencing; cyp51a.
• Online Access: http://jmsh.ac.in/index.php?option=com_k2&view=item&id=113:detection-of-azole-drug-resistance-of-the-aspergillus-species-cyp51a-gene-by-pcr&Itemid=88
• ISSN: 2394-9481; 2394-949X

Background: Allergic bronchopulmonary aspergillosis (ABPA) is a hypersensitivity pulmonary disease occurring in individuals with asthma or cystic fibrosis. In these patients, it is characterized by transient pulmonary infiltrates, reversible airway obstruction, eosinophilia, and evidence of hypersensitivity to the Aspergillus fumigatus. The aim of this study was standardization of antifungal drug susceptibility testing genotypic methods. Materials and Methods: This prospective and experimental study was carried out at the Department of Microbiology and Central Research Laboratory, Mahatma Gandhi Mission Medical College and Hospital, Navi Mumbai, and gene sequencing was carried out at Eurofins Laboratory. A molecular method for standardization of drug sensitivity testing for azole drug was done using Cyp51A gene and mutation M220 and L98H. The American Type Culture Collection control strain of Aspergillus oryzae, Aspergillus niger, and Aspergillus brasiliensis was obtained from Microbiologics Inc., USA. Results: Azole drug resistance by polymerase chain reaction (PCR) revealed 48/60 (80%), and azole drug resistance showed L98H and M220 genes in 32/60 (60%). Gene sequencing of Aspergillus species in patient samples revealed M220 and L98H mutation in cyp51A gene of Aspergillus Species. Conclusions: Azole drug resistance by PCR showed L98H and M220 in cyp51A gene. Gene sequencing showed similar resistance by PCR.