scholarly journals New Commercially Available IgG Kits and Time-Resolved Fluorometric IgE Assay for Diagnosis of Allergic Bronchopulmonary Aspergillosis in Patients with Cystic Fibrosis

2015 ◽  
Vol 23 (3) ◽  
pp. 196-203 ◽  
Author(s):  
Coralie Barrera ◽  
Bénédicte Richaud-Thiriez ◽  
Steffi Rocchi ◽  
Bénédicte Rognon ◽  
Sandrine Roussel ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Aspergillus Fumigatus ◽  
Allergic Bronchopulmonary Aspergillosis ◽  
Specific Ige ◽  
Enzyme Linked Immunosorbent Assay ◽  
Time Resolved ◽  
Content Type ◽  
Elisa Kit ◽  
Fungal Allergens ◽  
Specific Igg

ABSTRACTAllergic bronchopulmonary aspergillosis (ABPA) is difficult to diagnose; diagnosis relies on clinical, radiological, pathological, and serological criteria. Our aim was to assess the performance of two new commercially available kits and a new in-house assay: anAspergillus fumigatusenzyme-linked immunosorbent assay (ELISA) IgG kit (Bordier Affinity Products), anAspergillusWestern blotting IgG kit (LDBio Diagnostics), and a new in-house time-resolved fluorometric IgE assay (dissociation-enhanced lanthanide fluorescent immunoassay, or DELFIA) using recombinant proteins from anAspergillussp. recently developed by our laboratory for ABPA diagnosis in a retrospective study that included 26 cystic fibrosis patients.Aspergillus fumigatus-specific IgG levels measured by a commercial ELISA kit were in accordance with the level of precipitins currently used in our lab. The ELISA kit could accelerate and help standardize ABPA diagnosis.Aspergillus fumigatus-specific IgE levels measured by ImmunoCAP (Phadia) withA. fumigatusM3 antigen and by DELFIA with a purified protein extract ofA. fumigatuswere significantly correlated (P< 10−6). The results with recombinant antigens glucose-6-phosphate isomerase and mannitol-1-phosphate dehydrogenase were encouraging but must be confirmed with sera from more patients. The DELFIA is an effective tool that can detect specific IgE against more fungal allergens than can be detected with other commercially available tests.

scholarly journals Recombinant Allergens Combined with Biological Markers in the Diagnosis of Allergic Bronchopulmonary Aspergillosis in Cystic Fibrosis Patients

2010 ◽  
Vol 17 (9) ◽  
pp. 1330-1336 ◽  
Author(s):  
Hélène Fricker-Hidalgo ◽  
Bérangère Coltey ◽  
Catherine Llerena ◽  
Jean-Charles Renversez ◽  
Renée Grillot ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Retrospective Study ◽  
Biological Markers ◽  
Allergic Bronchopulmonary Aspergillosis ◽  
Specific Ige ◽  
Added Value ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Total Ige ◽  
Specific Igg

ABSTRACT Allergic bronchopulmonary aspergillosis (ABPA) is a frequent complication in cystic fibrosis patients. The diagnosis remains difficult and requires a combination of clinical, radiological, biological, and mycological criteria. The aim of this study was to analyze the added value of two recombinant antigens, rAspf4 and rAspf6, associated with the detection of specific IgG; precipitins; total IgE; and Aspergillus fumigatus in sputum for the diagnosis of ABPA. In a retrospective study, we determined the specific IgE responses to these recombinants in 133 sera of 65 cystic fibrosis patients. We selected an average of five serum samples from each of the 17 patients with ABPA (13 proven and 4 probable ABPA) and from 3 patients with Aspergillus bronchitis and rhinosinusitis. One serum sample for the 45 patients without ABPA was tested. The sensitivity of specific IgE detection against rAspf4 calculated per patient (92.3%) was significantly higher (P < 0.05) than that of rAspf6 (53.8%). When rAspf4 IgE detection was associated with anti-Aspergillus IgG enzyme-linked immunosorbent assay (ELISA) and precipitin detection, the sensitivity rose to 100%. The specificities of rAspf4 and rAspf6 IgE detection were 93.7% and 91.6%, respectively. Other diagnostic criteria had slightly lower specificities (87.5% for anti-Aspergillus IgG ELISA, 89.6% for precipitins, 84.4% for total IgE, and 85.0% for positive A. fumigatus culture in sputum). In conclusion, this retrospective study showed the relevance of rAspf4 IgE detection, in combination with other biological markers (Aspergillus IgG ELISA, precipitins, and total IgE), for improving the biological diagnosis of ABPA.

scholarly journals BASOPHILE ACTIVATION TEST FOR THE DIAGNOSTICS OF FUNGAL SENSITIZATION IN THE PATIENTS WITH CYSTIC FIBROSIS

2019 ◽  
Vol 21 (5) ◽  
pp. 919-928
Author(s):  
Ya. I. Kozlova ◽  
E. V. Frolova ◽  
A. E. Uchevatkina ◽  
L. V. Filippova ◽  
O. V. Aak ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Stimulation Index ◽  
Allergic Bronchopulmonary Aspergillosis ◽  
Basophil Activation Test ◽  
Specific Ige ◽  
Basophil Activation ◽  
Total Ige ◽  
Activation Test ◽  
Fungal Allergens ◽  
Fungal Sensitization

Aspergillus fumigatus colonization in the patients with cystic fibrosis (CF) may cause sensitization against A. fumigatus and/or allergic bronchopulmonary aspergillosis (ABPA), which significantly worsens the course of underlying disease. At the present time, new diagnostic tests are searched for detection of fungal sensitization in these patients. The aim of this work was to evaluate an opportunity of application of basophile activation test with A. fumigatus allergen in vitro using flow cytometry, aiming for identification of fungal sensitization in the CF patients. The study included 190 patients with CF aged 1 to 37 years. All the patients underwent common allergy screening (skin tests with fungal allergens, determination of serum levels of total IgE and specific IgE for the fungal allergens), and mycological examination (microscopy and culture of respiratory substrates). Computed tomography of the chest was performed upon clinical indications. The basophil activation test with the A. fumigatus allergen was performed in 10 CF patients with ABPA, and 10 CF patients without ABPA, in addition to the standard allergological examination. Frequency of sensitization to A. fumigatus in the patients with cystic fibrosis was 27%, the incidence of allergic bronchopulmonary aspergillosis was 5.7%. The number of eosinophils, total IgE and specific IgE levels in CF patients with ABPA were significantly higher than in CF patients without ABPA. In blood of the ABPA patients we have identified 68.5 (52.5-81.5%) of basophilic leukocytes activated by A. fumigatus allergen, with a stimulation index of 17.07 (10.30-27.70). In appropriate comparison group, the stimulation index did not exceed 1.5 (p = 0.000). Direct positive correlation between the levels of specific IgE to A. fumigatus and the number of basophils activated by A. fumigatus allergens was revealed (r = 0.77; р < 0.05). FVC values and the body mass index in CF patients with ABPA were significantly lower when compared with the patients without fungal sensitization. Introduction of the basophil activation test, along with standard techniques, may enable a more differentiated assessment of ABPA development in CF patients. Timely detection of associations between A. fumigatus sensitization and clinical status of CF patients will facilitate early and effective administration of specific therapy.

scholarly journals High Prevalence of Azole-Resistant Aspergillus fumigatus in Adults with Cystic Fibrosis Exposed to Itraconazole

2011 ◽  
Vol 56 (2) ◽  
pp. 869-874 ◽  
Author(s):  
Pierre-Régis Burgel ◽  
Marie-Thérèse Baixench ◽  
Michaël Amsellem ◽  
Etienne Audureau ◽  
Jeanne Chapron ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Aspergillus Fumigatus ◽  
Allergic Bronchopulmonary Aspergillosis ◽  
Previous Treatment ◽  
University Hospital ◽  
Clinical Implications ◽  
Content Type ◽  
Microdilution Method ◽  
Broth Microdilution Method ◽  
High Prevalence

ABSTRACTAspergillus fumigatusis the most frequent fungus found in the sputum of cystic fibrosis (CF) subjects. Itraconazole is prescribed for allergic bronchopulmonary aspergillosis (ABPA) orAspergillusbronchitis in CF subjects. We hypothesized thatA. fumigatusisolates in the sputum of CF subjects with previous exposure to itraconazole was associated with higher prevalence of azole resistance. From June 2010 to April 2011, sputum samples from adult CF subjects at Cochin University Hospital (France) were examined systematically for the detection ofA. fumigatus. MICs ofA. fumigatusisolates against azoles were screened using Etest, and reduced susceptibility to azoles was confirmed using the CLSI broth microdilution method.A. fumigatuswas isolated from the sputum of 131/249 (52.6%) adult CF subjects, and 47/131 (35.9%) subjects had received previous treatment with itraconazole. ReducedA. fumigatussusceptibility to itraconazole (MIC, ≥2 mg/liter) was confirmed in 6/131 (4.6%) subjects. All 6 isolates also had reduced susceptibility to posaconazole (MIC, ≥0.5 mg/liter), and 3/6 isolates had reduced susceptibility to voriconazole (MIC, ≥2 mg/liter). Mutations in thecyp51Agene were detected at positions previously implicated to cause resistance in 5 isolates. Azole-resistantA. fumigatusisolates were found in 5/25 (20%) subjects exposed to itraconazole within the previous 3 years. High rates of azole-resistantA. fumigatusisolates were present in adult CF subjects and were associated with recent itraconazole exposure. Although the clinical implications of these findings will require further studies, the cautious use of itraconazole in adult CF subjects can be recommended.

scholarly journals Aspergillus fumigatus In-Host HOG Pathway Mutation for Cystic Fibrosis Lung Microenvironment Persistence

mBio ◽  
2021 ◽  
Vol 12 (4) ◽  
Author(s):  
Brandon S. Ross ◽  
Lotus A. Lofgren ◽  
Alix Ashare ◽  
Jason E. Stajich ◽  
Robert A. Cramer
Keyword(s):  
Cystic Fibrosis ◽  
Aspergillus Fumigatus ◽  
Disease Progression ◽  
Allergic Bronchopulmonary Aspergillosis ◽  
Clinical Manifestations ◽  
Cystic Fibrosis Lung ◽  
Hog Pathway ◽  
Content Type

Aspergillus fumigatus infection causes a spectrum of clinical manifestations. For individuals with cystic fibrosis (CF), allergic bronchopulmonary aspergillosis (ABPA) is an established complication, but there is a growing appreciation for A. fumigatus airway persistence in CF disease progression.

scholarly journals Prospective Evaluation of Aspergillus fumigatus-Specific IgG in Patients With Cystic Fibrosis

2021 ◽  
Vol 10 ◽  
Author(s):  
Patience Eschenhagen ◽  
Claudia Grehn ◽  
Carsten Schwarz
Keyword(s):  
Cystic Fibrosis ◽  
Aspergillus Fumigatus ◽  
Sensitivity And Specificity ◽  
Statistical Significance ◽  
Allergic Bronchopulmonary Aspergillosis ◽  
Control Group ◽  
Opportunistic Fungi ◽  
Significant Difference ◽  
Specific Igg

BackgroundIn Cystic Fibrosis (CF), the airways are often colonized by opportunistic fungi. The most frequently detected mold is Aspergillus fumigatus (Af). Af diseases are associated with significant morbidity and mortality. The most common clinical picture caused by Af is allergic bronchopulmonary aspergillosis (ABPA), triggered by an immunological reaction against Af. Af bronchitis and invasive aspergillosis rarely occur in CF as a result of spore colonization and germination. Since pulmonary mycoses and exacerbations by other pathogens overlap in clinical, radiological, and immunological characteristics, diagnosis still remains a challenge. The search for reliable, widely available biomarkers for Af diseases is therefore still an important task today.ObjectivesAf-specific IgG m3 is broadly available. Sensitivity and specificity data are contradictory and differ depending on the study population. In our prospective study on pulmonary Af diseases in CF, we determined specific IgG m3 in order to test its suitability as a biomarker for acute Af diseases and as a follow-up parameter.MethodsIn this prospective single center study, 109 patients with CF were screened from 2016 to 2019 for Af-associated diseases. According to diagnostic criteria, they were divided into four groups (control, bronchitis, ABPA, pneumonia). The groups were compared with respect to the level of Af-specific IgG (ImmunoCAP Gm3). We performed a receiver operating characteristic (ROC) curve analysis to determine cut-off, sensitivity and specificity. Twenty-one patients could be enrolled for a follow-up examination.ResultsOf the 109 patients, 36 were classified as acute Af-disease (Af bronchitis, ABPA, Af pneumonia). Of these, 21 patients completed follow up-screening. The median Af-specific Gm3 was higher in the acute Af-disease groups. There was a significant difference in Af-specific IgG m3 compared to the control group without acute Af-disease. Overall, there was a large interindividual distribution of Gm3. A cut-off value of 78.05 mg/L for Gm3 was calculated to discriminate controls and patients with ABPA/pneumonia with a specificity of 75% and a sensitivity of 74.6%. The follow up examination of 21 patients showed a decrease of Gm3 in most patients without statistical significance due to the small number of follow up patients.ConclusionAf specific IgG may be a useful biomarker for acute ABPA and Af pneumonia, but not for Af bronchitis in CF. However, due to the large interindividual variability of Gm3, it should only be interpreted alongside other biomarkers. Therefore, due to its broad availability, it could be suitable as a biomarker for ABPA and Af pneumonia in CF, if the results can be supported by a larger multicenter cohort.

scholarly journals Features of fungal sensitization in patients with asthma and cystic fibrosis: the results of a prospective study

2018 ◽  
Vol 15 (2) ◽  
pp. 11-16
Author(s):  
Y I Kozlova ◽  
E V Frolova ◽  
Y V Borzova ◽  
A V Sobolev ◽  
E V Burygina ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Allergic Bronchopulmonary Aspergillosis ◽  
Specific Ige ◽  
Test System ◽  
Candida Spp ◽  
Ige Antibodies ◽  
Fungal Allergens ◽  
Patients At Risk ◽  
Fungal Sensitization

Background. To estimate the frequency of fungal sensitization and allergic bronchopulmonary aspergillosis in patients with asthma or cystic fibrosis. Methods. The study included 205 patients with asthma and 190 patients with cystic fibrosis. For detection of fungal sensitization specific IgE antibodies in the blood serum with «AllergoELISA-specific IgE» test system and biotinylated fungal allergens were determined. Mycological examination included microscopy and cultural study of respiratory biomaterials. According to the indications, a CAT scan of the chest was performed. Results. The incidence of fungal sensitization in patients with asthma was 35,6%, CI [29,1%; 42,6%], in patients with cystic fibrosis - 56,8% [49,5%; 64%]. In patients with severe asthma the main fungal allergens were Alternaria spp. (70%) and Aspergillus spp. (60%); in patients with cystic fibrosis: Candida spp. - 73%, Alternaria spp. - 34%, Aspergillus spp. - 27%. The incidence of allergic bronchopulmonary aspergillosis (ABPA) in patients with asthma was 5,3%, in patients with cystic fibrosis - 5,7%. Conclusion Early detection of ABPA in patients with asthma and cystic fibrosis is very important as it changes the therapeutic measures of patients’ management and prevents progression of bronchiectasis and respiratory failure. Determination of specific Aspergillus spp. IgE antibodies in vitro is a necessary stage of diagnosis of ABPA in patients at risk.

scholarly journals Eosinophil Deficiency Compromises Lung Defense against Aspergillus fumigatus

2013 ◽  
Vol 82 (3) ◽  
pp. 1315-1325 ◽  
Author(s):  
Lauren M. Lilly ◽  
Michaella Scopel ◽  
Michael P. Nelson ◽  
Ashley R. Burg ◽  
Chad W. Dunaway ◽  
...  
Keyword(s):  
Aspergillus Fumigatus ◽  
Allergic Bronchopulmonary Aspergillosis ◽  
Cell Contact ◽  
Mrna Levels ◽  
Colony Stimulating Factor ◽  
Content Type ◽  
Eosinophil Peroxidase ◽  
Stimulating Factor ◽  
Deficient Mice

ABSTRACTExposure to the moldAspergillus fumigatusmay result in allergic bronchopulmonary aspergillosis, chronic necrotizing pulmonary aspergillosis, or invasive aspergillosis (IA), depending on the host's immune status. Neutrophil deficiency is the predominant risk factor for the development of IA, the most life-threatening condition associated withA. fumigatusexposure. Here we demonstrate that in addition to neutrophils, eosinophils are an important contributor to the clearance ofA. fumigatusfrom the lung. AcuteA. fumigatuschallenge in normal mice induced the recruitment of CD11b+Siglec F+Ly-6GloLy-6CnegCCR3+eosinophils to the lungs, which was accompanied by an increase in lungEpx(eosinophil peroxidase) mRNA levels. Mice deficient in the transcription factor dblGATA1, which exhibit a selective deficiency in eosinophils, demonstrated impairedA. fumigatusclearance and evidence of germinating organisms in the lung. Higher burden correlated with lower mRNA expression ofEpx(eosinophil peroxidase) andPrg2(major basic protein) as well as lower interleukin 1β (IL-1β), IL-6, IL-17A, granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), and CXCL1 levels. However, examination of lung inflammatory cell populations failed to demonstrate defects in monocyte/macrophage, dendritic cell, or neutrophil recruitment in dblGATA1-deficient mice, suggesting that the absence of eosinophils in dlbGATA1-deficient mice was the sole cause of impaired lung clearance. We show that eosinophils generated from bone marrow have potent killing activity againstA. fumigtausin vitro, which does not require cell contact and can be recapitulated by eosinophil whole-cell lysates. Collectively, our data support a role for eosinophils in the lung response afterA. fumigatusexposure.

scholarly journals Development of a Recombinant Protein-Based Enzyme-Linked Immunosorbent Assay for Diagnosis of Mycoplasma bovis Infection in Cattle

2013 ◽  
Vol 21 (2) ◽  
pp. 196-202 ◽  
Author(s):  
Nadeeka K. Wawegama ◽  
Glenn F. Browning ◽  
Anna Kanci ◽  
Marc S. Marenda ◽  
Philip F. Markham
Keyword(s):  
Enzyme Linked Immunosorbent Assay ◽  
Mycoplasma Bovis ◽  
Western Blots ◽  
Content Type ◽  
Amino Terminal ◽  
Igm Elisa ◽  
Specific Igg ◽  
Lipase A ◽  
Strong Immunoreactivity ◽  
Gst Fusion Proteins

ABSTRACTMycoplasma boviscauses a range of diseases in cattle, including mastitis, arthritis, and pneumonia. However, accurate serological diagnosis of infection remains problematic. The studies described here aimed to identify an antigen that might be used to develop a more specific and sensitive diagnostic assay. A 226-kDa immunogenic protein was consistently detected in Western blots by antibodies in sera from calves experimentally infected withM. bovis. This protein was shown to be a membrane protein with lipase activity and was named mycoplasma immunogenic lipase A (MilA). Different regions of MilA were expressed inEscherichia colias glutathioneS-transferase (GST) fusion proteins and recombinant products from the amino-terminal end shown to have strong immunoreactivity withM. bovis-specific bovine sera. The most immunoreactive fusion protein, GST-MilA-ab, was used to develop indirect IgM and IgG enzyme-linked immunosorbent assays (ELISAs). The IgM ELISA detectedM. bovis-specific IgM antibody 2 weeks after infection with 97.1% sensitivity and had a specificity of 63.3%, while the IgG ELISA detectedM. bovis-specific IgG 3 weeks after infection with 92.86% sensitivity and had a specificity of 98.7%, demonstrating that the IgG ELISA has potential for use as a sensitive and specific assay for detecting infection in cattle.

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