MOLECULAR DETECTION AND CLUSTER ANALYSIS OF VACUOLAR SERINE PROTEASE GENE IN PENICILLIUM ISOLATED FROM HOSPITAL INDOOR AIR IN YOGYAKARTA, INDONESIA

摘要

Penicillium are commonly found in indoor air and some of them are known as allergenic fungi. Vacuolar serine protease (VSP) was reported as major allergens of several Penicillium species. Existence of allergenic fungi is considered as a risk factor for inhabitants inside the room, especially in hospital. So far, no studies are available about hospital airborne fungi in Indonesia. Therefore, this study was conducted to investigate fungal isolates obtained in hospital indoor air and then focused on detection as well as cluster analysis of VSP allergenic Penicillium. All fungal genera obtained were identified in genus level based on morphological characteristics. Fungal group belong to Penicillium were then collected for further molecular detection based on Polymerized Chain Reaction (PCR) amplification of VSP gene using new degenerate primer design (VP1 and VP2) and continued with species identification based on (3-tubulin gene. Cluster analysis was exhibited by phylogenetic relationship among VSP allergenic Penicillium isolates based on nucleotide sequence of VSP gene. The result showed that genus of Penicillium was found as dominant airborne fungi from several hospital rooms. Moreover, presence of VSP gene was detected in P.citrinum, P. steckii,and P. copticola. Interestingly, the existing of VSP gene in P. steckii and P. copticola is the first time reported especially in Indonesia. Furthermore, phylogenetic tree constructed as cluster analysis showed that VSP gene of the same Penicillium species was grouped in one cluster. It indicates that gene encoding VSP allergen might be classified as species-specific.