Calcineurin-dependent galactomannan release in Aspergillus fumigatus

Item does not contain fulltextThe galactomannan assay to diagnose invasive aspergillosis is recommended and clinically utilized, yet the mechanism of galactomannan release from Aspergillus fumigatus is unknown. We used an A. fumigatus strain lacking calcineurin A (cnaA), already shown to be critically important for pathogenicity, to evaluate galactomannan kinetics. During the logarithmic growth phase when glucose was consumed, beta-D-galactofuranoside (galf)-antigens were released in the culture. However, after glucose became limited, GM release further increased in the supernatants of the wild type strain while there was no further increase of GM release in the DeltacnaA strain. beta-Galactofuranosidase activity was also decreased in the DeltacnaA mutant, and the amount of galf-antigen in the cell wall fraction of the DeltacnaA mutant was approximately ten-fold higher. This suggests the possibility that the antigen is unable to be released due to a cell wall abnormality. This and previous work suggest a dynamic calcineurin-dependent cell wall during periods of growth, with galactomannan release from the cell wall possibly calcineurin-dependent and reflected in the decreased GM release and greatly increased cell wall fraction of galf in the DeltacnaA mutant

Paradoxical increase in circulating Aspergillus antigen during treatment with caspofungin in a patient with pulmonary aspergillosis.

Contains fulltext : 50418.pdf (publisher's version ) (Open Access)A paradoxical increase in circulating Aspergillus antigen was observed during treatment with caspofungin in a patient with proven invasive aspergillosis. With the exception of treatment with the echinocandin, no other factors were found that might explain this clinical observation, which was supported by experiments done in vitro

Improved Detection of Circulating Aspergillus Antigen by Use of a Modified Pretreatment Procedure▿

Detection of circulating galactofuranose (galf) antigens, including galactomannan (GM), by the Platelia Aspergillus (PA) enzyme-linked immunosorbent assay (ELISA) is an important tool in the early diagnosis of invasive aspergillosis (IA). We used a modified pretreatment technique (MT) on consecutive negative PA ELISA plasma samples from IA patients in order to improve the detection of the 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. The PA ELISA reactivity of pretreated samples was determined before (conventional technique [CT]) and after (MT) filtration using a Microcon filter with a 50-kDa cutoff (Millipore). For the MT, the sensitivity of the PA ELISA increased from 42.9% (CT) to 78.6% (MT) using a cutoff 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

Bifidobacterial Lipoglycan as a New Cause for False-Positive Platelia Aspergillus Enzyme-Linked Immunosorbent Assay Reactivity

We previously hypothesized that a lipoglycan of Bifidobacterium bifidum subsp. pennsylvanicum cross-reacts with the Platelia Aspergillus (PA) enzyme-linked immunosorbent assay (ELISA) based on the presence of galactofuranosyl epitopes in the cell wall (M. A. S. H. Mennink-Kersten, R. R. Klont, A. Warris, H. J. M. Op den Camp, and P. E. Verweij, Lancet 363:325-327, 2004). We tested this hypothesis by testing bacterial suspensions of different bifidobacterial species and other gram-positive and -negative bacteria with the PA ELISA, which is used to detect circulating galactomannan for the serodiagnosis of invasive aspergillosis. Furthermore, neonatal fecal samples were enumerated for bifidobacteria by fluorescence in situ hybridization (FISH) and tested for PA ELISA reactivity. All bifidobacteria, except B. infantis and B. adolescentis, showed reactivity 6- to 600-fold higher compared to the controls (i.e., Micrococcus luteus and Propionibacterium freudenreichii, which contain a cell wall lipomannan). Eggerthella lenta showed a 25-fold-higher reactivity. ELISA reactivity was clearly shown to be associated with bacterial lipoglycans containing a β-1,5-galactofuranosyl chain. All neonatal feces showed PA ELISA reactivity and associated numbers of bifidobacteria. Since high concentrations of bifidobacteria are present in the human gut, these bacteria or excreted lipoglycan may cause false serum PA ELISA reactivity in selected patient groups, especially neonates

Value of Candida serum markers in patients with invasive candidiasis after myeloablative chemotherapy

Invasive Candida infections are associated with a significant morbidity and mortality. Detection of circulating biomarkers has been shown to precede conventional diagnostic methods, which is important in improving outcome. We investigated the performance of multiple biomarkers using Candida antigen and anti-Candida antibody detection systems of Platelia and Serion and beta-d-glucan detection in serial serum samples from patients, treated for leukemia, with invasive candidiasis. The performance of single assays and combined detection appeared different for patients with 1 or more episodes of neutropenia and is therefore related to the phase of therapy for the underlying leukemia of the patient. These new insights may help to optimize the diagnostic strategies for the diagnosis of invasive candidiasis

Utility of Aspergillus antigen detection in specimens other than serum specimens.

Item does not contain fulltextThe detection of circulating galactomannan in serum is an important tool for the early diagnosis of invasive aspergillosis. A commercial enzyme-linked immunosorbent assay (Platelia Aspergillus; BioRad) was shown to be both highly sensitive and specific for detection of galactomannan in serum samples. Despite the fact that this assay is validated for serum samples, specimens of other body fluids are increasingly used for detection of galactomannan, including urine, bronchoalveolar lavage fluid, and cerebrospinal fluid. Review of the literature shows that galactomannan can be detected in each of these samples from patients with invasive aspergillosis with higher sensitivity than is the case with culture, as well as early in the course of infection. However, the evidence thus far is based on case reports--predominantly retrospective studies--that often include heterogeneous patient populations and limited numbers of cases of proven infection. Clearly, well-designed prospective studies with systematic sampling and use of consensus case definitions are needed to compare the performance of antigen detection in samples other than serum specimens with that in serum specimens