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Plasma ��-d-glucan was measured with the ��-glucan test WAKO (Wako Pure Chemical Industries, Tokyo, Japan). Plasma samples were collected prior to the collection of BAL or IS samples and the initiation of treatment. The assay was performed at our institution on the same day that the plasma was obtained. Categorical variables were compared using the Fisher exact test. Continuous variables were compared using the Kruskal�CWallis test or the Mann�CWhitney U-test. ��-d-glucan values under the detection limit of 4.3?pg/mL CYTH4 were considered to be 4.3?pg/mL. Receiver-operating characteristic (ROC) curves for the ��-d-glucan levels and copy numbers were constructed, and their optimal cut-off values were determined with the maximum Youden index. We compared the area under the curve (AUC) of the two ROC curves using the method described by Delong et?al. [18]. A p value check details PCP, colonization, non-colonization or undetermined was made in 11, 42, 15, 60 and 19 patients, respectively. BAL samples were obtained from 7, 10, 6, 12 and five of these patients, respectively. All definite patients received anti-PCP treatment. The final diagnoses of the colonized patients included bacterial pneumonia (n?=?4), interstitial pneumonia (n?=?3), invasive aspergillosis (n?=?2), atypical BIBF1120 pneumonia (n?=?1), acute respiratory distress syndrome (ARDS) due to sepsis (n?=?1), tuberculosis (n?=?1), pulmonary embolism (n?=?1), cryptogenic organizing pneumonia (n?=?1), and pulmonary alveolar proteinosis (n?=?1). Among the non-colonized patients, the final diagnoses included interstitial pneumonia (n?=?18), bacterial pneumonia (n?=?10), ARDS due to sepsis (n?=?9), drug-induced pneumonia (n?=?5), pulmonary haemorrhage (n?=?5), viral pneumonia (n?=?3), tuberculosis (n?=?2), pulmonary oedema (n?=?1), invasive aspergillosis (n?=?1), radiation pneumonitis (n?=?1), lymphangitis carcinomatosa (n?=?1), cryptogenic organizing pneumonia (n?=?1), and unknown (n?=?3). Table?1 shows the patient demographics and characteristics. Three definite patients and two probable patients had HIV infection, whereas none of the colonized or non-colonized patients did. An analysis of laboratory test results revealed significant differences in thrombocyte, C-reactive protein, blood urea nitrogen and total bilirubin levels between the four groups. The qPCR and ��-d-glucan assay results are presented in Fig.?1. All definite patients and 40 of the 42 probable patients were qPCR-positive. Copy numbers were significantly higher in the definite group (median 460?000?copies/mL, range 1400�C15?000?000) than in the colonized group (median 180, range 10�C15?000; p?