Comparison of Real-Time Aspergillus PCR with Platelia™AspergillusEIA in broncho-alveolar lavage fluids for the diagnosis of invasive aspergillosis in neutropenic and non-neutropenic patients

Abstract

Objectives. Because of low sensitivity of fungal cultures and lack of standardization of Aspergillus PCR, laboratory diagnosis of invasive aspergillosis still relies on galactomannan (GM) detection in broncho-alveolar lavage (BAL) fluids and serum. The aim of this study was double: first, to assess the place of a 18S rRNA Aspergillus real-time PCR test performed in BAL fluid for the diagnosis of invasive aspergillosis (IA) in neutro- and non-neutropenic patients in comparison with GM detection; secondly, to evaluate the use of three different GM cut-off values. Materials and methods. A total of 111 neutropenic and non-neutropenic patients hospitalized at the University hospital of Liège from March to October 2012 with suspicion of IA were included in the study. A total of 138 broncho-alveolar lavage fluids were evaluated by three laboratory diagnostic methods: 1/ culture on Sabouraud agar slants with antibiotics (bioMérieux, France) incubated at 28°C for 28 days; 2/ GM detection (Platelia ™Aspergillus EIA, Biorad) using GM index cut-off values at 0.5, 0.8 and 1, performed three times a week; 3/ a real-time Aspergillus PCR assay performed daily and targeting the 18S rRNA genes by using an in-house method. Clinical, radiological and microbiological data were reviewed for classification of patients. Results. Nine patients developed probable or possible IA. The sensitivity/specificity/positive (VPP) and negative (NPV) predictive values (%) for culture, PCR, and GM using 0,5 as cut-off value were respectively 41/100/100/94, 58/97/70/96, and 91/83/34/99. The use of 0,8 and 1 as GM index cut-off values increased the specificity to 89 and 92% respectively, and the VPP to 44 and 54%. PCR had a better turn-around time and allowed the detection of Aspergillus colonisation. Conclusion: GM detection in BAL fluids using a cut-off value of 1 was the most efficient laboratory test for the diagnosis of IA in neutropenic and non-neutropenic patients. Despite a lower sensitivity, PCR had a better VPP, and allowed the detection of culture-negative Aspergillus colonisations. A shorter turnaround time (TAT) due to daily practice of PCR tests may reduce the time-to-treatment up to 24 hours