POLYMERASE CHAIN-REACTION AMPLIFICATION OF ASP-F-I AND ALKALINE PROTEASE GENES FROM FUNGUS BALLS - CLINICAL-APPLICATION IN PULMONARY ASPERGILLOSIS

AspfI(18 kDa) and alkaline protease (33 kDa) are the 2 major antigens which are derived from Aspergillus (A,)fumigatus and have been implica ted as possible virulence factors in the pathogenesis of Aspergillus-i nduced diseases. We attempted to detect fragments of genes encoding bo th proteins from fungus balls obtained at surgery or autopsy by polyme rase chain reaction (PCR) amplification and then used PCR to test clin ical samples, Frozen-stored fungus ball samples from a patient with ac ute myeloid leukemia complicated by Aspergillus pneumonia and from a p atient with pulmonary aspergilloma were studied, We successfully ampli fied a 315 bp PCR product, the target sequence for AspfI, and a 747 bp PCR product as a target sequence for alkaline protease (ALP) in both cases, In addition, 13 clinical samples including sputum specimens fro m patients with pulmonary aspergillosis were also examined, PCR analys is for the AspfI (ALP) gene in clinical samples showed positive result s in 5/10 (6/10) patients with pulmonary aspergilloma and in 3/3 (1/3) patients with invasive pulmonary aspergillosis. Culture data on A, fu migatus revealed positive results in 3/9 patients with pulmonary asper gilloma and in 2/3 patients with invasive pulmonary aspergillosis, Thi s method can be used to recognize the involvement of A, fumigatus in v arious clinical settings where conventional culture results are not re adily available.