Improved Detection of Circulating Aspergillus Antigen Using a Modified Pre-treatment Procedure

Author:

Mennink-Kersten MA, Ruegebrink D, Klont RR, Warris A, Blijlevens NM, Donnelly JP, Verweij PE

Date: 14 March 2008

Abstract:

Detection of circulating galactofuranose (galf)-antigens, including galactomannan (GM), by the Platelia Aspergillus (PA) ELISA is an important tool in the early diagnosis of invasive aspergillosis (IA). We used a modified pre-treatment technique (MT) on consecutive PA ELISA negative plasma samples from IA patients in order to improve the detection of fungal components present. Plasma samples (52) were collected from healthy donors and 174 plasma samples with a galactomannan index (GMI) below 0.5 were collected from 25 unclassifiable and 23 IA patients. PA ELISA reactivity of pre-treated samples was determined before (conventional technique, CT) and after filtration (MT) using a Microcon filter with a 50-kDa cut-off (Millipore). For the MT, the sensitivity of the PA ELISA increased from 42.9 % (CT) to 78.6 % (MT) using a cut-off for the GMI of 1.5 in the probable and proven group, whereas specificity slightly decreased from 98.7 % to 96.1 % in the control group. The 10-fold concentration step increased the GMI as high as 121-fold. The MT resulted not only in positive reactivity in samples that tested negative with the CT but also in the earlier detection of antigen by 2 to 17 days.

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