Respiratory Treatment and Prevention (Advances in Experimental Medicine and Biology)

2 Methods

The Bioethical Commission of the Medical Uni-
versity of Warsaw in Poland waived the require-
ment for ethical approval due to the retrospective
nature of this research. The prevalence ofAsper-
gillus spp. infection in patients hospitalized in
the the Infant Jesus Teaching Hospital in
Warsaw, Poland during the years 2013–2014
was retrospectively investigated. Routine myco-
logical tests revealed 200 isolates ofAspergillus
spp. from the respiratory tract, including 184 tra-
cheal aspirates and 16 bronchoalveolar lavages,
among 2000 clinical samples of fungi.

2.1 Culture and Identification

Strains were cultured on the Sabouraud agar with
chloramphenicol at 30C for 24–48 h and were
then identified by morphological characteristics
under a microscope. Microscopic features of
A. fumigatus consisted of typical columnar,
uniseriate conidial heads. Conidiophores are
short, smooth-walled, and have conical shaped
terminal vesicles which support a single row of
phialides on the upper two thirds of the vesicle.
Conidia are produced in basipetal succession,
forming long chains. Conidia are globose to

subglobose, green, and rough-walled to

echinulate (Andreoni et al. 2004 ).

2.2 Galactomannan (GM) Platelia

AspergillusEnzyme

Immunoassay (EIA)

The serum was taken for serological testing. All

collected serum samples were stored at 20 C

until use. The sandwich ELISA method was

employed to detect galactomannan in the serum

according to the manufacturer’s instruction

(Platelia Aspergillus protocol; BioRad, Marnes-

la-Coquette, France). Optical density was

measured spectrophotometrically with BioRad

Model PR5100 ELISA microplate reader. The

results were interpreted as based on the index

calculated from the measured optical density

using the 450 nm wavelength. A cut-off index

of 0.5 was considered positive.

3 Results

Two thousand clinical samples testing positive

for fungi, including 200 from the respiratory

tract, were analyzed within a 2-year period of

2013–2014. A. fumigatus was identified by

Fig. 1 Classification of invasive infections according to EORTC/MSG guidelines (de Pauw et al. 2008 ; Ascioglu
et al. 2002 )

Diagnosis of Invasive Pulmonary Aspergillosis 29