Performance of a commercial real-time PCR assay using the ABI 7300 instrument for the detection of Aspergillus and Pneumocystis DNA in bronchoalveolar lavage fluid samples from critical patient

C.F. Orsi*, W. Gennari, C. Venturelli, G. Bertazzoni, A. La Regina, M. Machetti, M. Pecorari, F. Rumpianesi, E. Blasi

Author address: 

(Modena, Genoa, IT)

Abstract: 

Objectives: To investigate the performance of the MycAssayTM Aspergillus (MycAspAssay) and of the MycAssayTM Pneumocystis (MycPCPAssay) in detecting Aspergillus (Asp) or P. jirovecii (Pj) DNA in BAL samples from critical patients, using ABI 7300 instrument. Methods: We enrolled 20 patients with or without fungal infections, according to the EORTC classification. In detail, seven patients were diagnosed with invasive aspergillosis (IA), eight patients with pulmonary infection by Pj (PCP) and five hospitalized patients with non-fungal infections (CTRL). DNA extracted by MycXtraTM (Myconostica) from BAL samples was amplified by real-time PCR by MycAspAssay and MycPCPAssay kits (Myconostica) using ABI 7300 instrument (Applied Biosystem). The real-time PCR data were compared with the results obtained with conventional diagnostic tests, namely culture, galactomannan (GM), immunofluorescence (IF), sequencing and Aspergillus spp. Q-PCR Alert Kit or P. jirovecii Alert kit (Nanogen). Results: All the seven patients with IA as well as one of eight patients belonging to the PCP group were MycAspAssay positive whereas all the CTRL-group patients (n = 5) provided negative PCR results. All the MycAspAssay positive results were confirmed either by GM-assay or Aspergillus spp. Q-PCR Alert Kit. Seven of the eight patients with PCP as well as four of the seven patients belonging to the IA group were MycPCPAssay positive, whereas all the CTRL-group patients provided negative PCR results. Six of the 7 MycPCPAssay positive results were in line with the IF, nested PCR and sequencing data. Conclusion: Although being considered as a preliminary step towards a validation of the MycAssayTM Aspergillus and MycAssayTM
2012

abstract No: 

P831

Full conference title: 

22nd European Congress of Clinical Microbiology and Infectious Diseases
    • ECCMID 22nd (2012)