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 ◽  
...  

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.

2015 ◽  
Vol 23 (3) ◽  
pp. 196-203 ◽  
Author(s):  
Coralie Barrera ◽  
Bénédicte Richaud-Thiriez ◽  
Steffi Rocchi ◽  
Bénédicte Rognon ◽  
Sandrine Roussel ◽  
...  

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.


2019 ◽  
Vol 21 (5) ◽  
pp. 919-928
Author(s):  
Ya. I. Kozlova ◽  
E. V. Frolova ◽  
A. E. Uchevatkina ◽  
L. V. Filippova ◽  
O. V. Aak ◽  
...  

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.


2021 ◽  
Author(s):  
Nasamon Wanlapakorn ◽  
Nungruthai Suntronwong ◽  
Harit Phowatthanasathian ◽  
Ritthideach Yorsang ◽  
Thanunrat Thongmee ◽  
...  

Abstract Limited data are available on the responses to heterologous vaccine regimens for SARS-CoV-2, especially among countries using inactivated and adenoviral-vectored vaccines. A total of 77 participants who received heterologous prime/booster-inactivated COVID-19 vaccine and adenoviral-vectored vaccine were enrolled in our study. There were two comparison groups vaccinated with the homologous CoronaVac (N = 79) and AZD1222 (N = 80) regimen. All sera samples were tested for SARS-CoV-2 spike receptor-binding-domain (RBD) IgG using a chemiluminescent microparticle immunoassay (CMIA). The neutralizing activity in a subset of serum samples was tested against the original Wuhan strain and variants of concern, B.1.1.7 and B.1.351, using an enzyme-linked immunosorbent assay (ELISA)-based surrogate virus neutralization test (sVNT). The CoronaVac followed by the AZD1222 vaccine induced higher levels of spike RBD-specific IgG than that of two-dose CoronaVac or AZD1222 vaccines (p < 0.001). Sera samples of the CoronaVac/AZD1222 vaccine recipients elicited higher neutralizing antibody activity against the original Wuhan and the B.1.351 strain than in the recipients of the two-dose CoronaVac or AZD1222. Following the inactivated CoronaVac/adenoviral-vectored (AZD1222) vaccination administered 14–72 days apart, participants receiving the heterologous vaccine regimen had higher spike RBD-specific IgG and neutralizing activities than the homologous CoronaVac vaccine recipients.


2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Edurne Peña-Mendizabal ◽  
Sergi Morais ◽  
Ángel Maquieira

Abstract New antigens deriving from -lloyl and -llanyl, major and minor determinants, respectively, were produced for β-lactam antibiotics cefuroxime, cefotaxime, ceftriaxone, meropenem and aztreonam. Twenty β-lactam antigens were produced using human serum albumin and histone H1 as carrier proteins. Antigens were tested by multiplex in vitro immunoassays and evaluated based on the detection of specific IgG and IgE in the serum samples. Both major and minor determinants were appropriate antigens for detecting specific anti-β-lactam IgG in immunised rabbit sera. In a cohort of 37 allergic patients, we observed that only the minor determinants (-llanyl antigens) were suitable for determining specific anti-β-lactam IgE antibodies with high sensitivity (< 0.01 IU/mL; 24 ng/L) and specificity (100%). These findings reveal that not only the haptenisation of β-lactam antibiotics renders improved molecular recognition events when the 4-member β-lactam ring remains unmodified, but also may contribute to develop promising minor antigens suitable for detecting specific IgE-mediated allergic reactions. This will facilitate the development of sensitive and selective multiplexed in vitro tests for drug-allergy diagnoses to antibiotics cephalosporin, carbapenem and monobactam.


1980 ◽  
Vol 89 (3_suppl) ◽  
pp. 133-137 ◽  
Author(s):  
John L. Sloyer ◽  
John H. Ploussard ◽  
Laurel J. Karr

IgE antibody directed against noncapsular antigens of mechanically disrupted Streptococcus pneumoniae, serotype 3 rough, was demonstrated in middle ear effusions (MEE) and serum of infants with and without prior evidence of pneumococcal otitis media with effusion (OME). The techniques employed included radioimmunoassay (RIA), passive skin testing, Prausnitz-Küstner (P/K), and enzyme-linked immunospecific assay (ELISA). Adsorption of MEE with ultrasonically disrupted crude pneumococcal antigen (CPA-U) resulted in a reduction of total IgE counts per minute and suggested bacteria-specific IgE antibody ranging from approximately 22 to 92% of the total IgE. The biological activity of the IgE antibody was confirmed by challenging skin passively sensitized with MEE IgE and CPA-U. Areas of induration appeared 20 minutes after challenge and continued to increase in size until 90 minutes. An ELISA procedure was developed as a tool to determine the nature of the antigen(s) and to determine the class(es) of antibody other than IgE. It appeared that CPA-U possesses free amino groups and that it can withstand the rigors of autoclaving. An analysis of 45 cord bloods revealed that high levels of IgG CPA-U antibody occur in this type of sample and that no correlation exists between the IgE and IgG levels. The mean IgG: IgE ratio, optical density at 420 nm (OD 420), for cord bloods was 2.49. In contrast, serum samples from nine infants without pneumococcal otitis media and from 14 infants with pneumococcal otitis media had lower levels of IgG antibody. There was no significant relationship between IgG and IgE OD 420 in infants who never had an episode of pneumococcal otitis media and the mean IgG: IgE was 1.88, whereas the ratio for those infants with pneumococcal otitis media was 1.56. In addition, there was a significant correlation between IgG and IgE levels in this latter group. The results suggest that it may be important to monitor the levels of at least these two classes of antibody to enhance our understanding of the pathogenesis and recovery from otitis media.


2015 ◽  
Vol 22 (11) ◽  
pp. 1154-1159 ◽  
Author(s):  
D. Goldblatt ◽  
C. Y. Tan ◽  
P. Burbidge ◽  
S. McElhiney ◽  
L. McLaughlin ◽  
...  

ABSTRACTThe pneumococcal enzyme-linked immunosorbent assay (ELISA) reference standard serum, lot 89SF, has been in use since 1990 and was replaced in 2013 with a new reference standard, 007sp, that is projected to be available for the next 25 years. 007sp was generated under an FDA-approved clinical protocol; 278 adult volunteers were immunized with the 23-valent unconjugated polysaccharide vaccine Pneumovax II, and a unit of blood was obtained twice from each immunized subject within 120 days following immunization. Pooled serum was prepared from the plasma of 262 subjects, filled at 6 ml per vial, and lyophilized. Five independent laboratories participated in bridging the serotype-specific IgG assignments for 89SF to the new reference standard, 007sp, to establish equivalent reference values for 13 pneumococcal capsular serotypes (1,3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, and 23F) by using the WHO reference ELISA. In a second study involving three laboratories, a similar protocol was used to assign weight-based IgG concentrations in micrograms per ml to 007sp of seven serotypes (8, 10A, 11A, 12F, 15B, 22F, and 33F) also present in the 23-valent pneumococcal unconjugated polysaccharide vaccine. In addition, the IgG assignments for a 12-member WHO quality control (QC) serum panel were also extended to cover these seven serotypes. Agreement was excellent, with a concordance correlation coefficient (rc) of >0.996 when each laboratory was compared to the assigned values for the 12 WHO QC serum samples. There are four remaining pneumococcal serotypes (2, 9N, 17F, and 20) found in Pneumovax II for which IgG assignments exist for 89SF and remain to be bridged.


2008 ◽  
Vol 82 (3) ◽  
pp. 251-254 ◽  
Author(s):  
P. Mesén-Ramírez ◽  
E. Abrahams-Sandí ◽  
K. Fernández-Quesada ◽  
P. Morera

AbstractAngiostrongylus costaricensis is the aetiological agent of human abdominal angiostrongyliasis, a parasitic disease reported from the United States to Argentina, with a widespread occurrence of the nematode throughout Central and South America. This study assesses the performance of A. costaricensis eggs as antigen in an enzyme-linked immunosorbent assay (ELISA), for the determination of parasite-specific IgG1 antibodies. The specificity and the sensitivity of the method were 87% and 90.5%, respectively. Through this test it was possible to demonstrate a sharp and early decline in IgG1 antibody in serum samples taken from patients with histopathological diagnosis of abdominal angiostrongyliasis at different time points after surgical treatment. The present work demonstrated the usefulness of the egg antigen in the development of a specific diagnostic test for abdominal angiostrongylosis.


Parasitology ◽  
2012 ◽  
Vol 139 (7) ◽  
pp. 956-961 ◽  
Author(s):  
A. L. R. GONÇALVES ◽  
D. S. NUNES ◽  
M. R. F. GONÇALVES-PIRES ◽  
M. T. UETA ◽  
J. M. COSTA-CRUZ

SUMMARYThe aim of this study was to use larval, parasitic female and egg antigens fromStrongyloides venezuelensisto detect parasite-specific IgG and immune complexes in human serum samples by enzyme-linked immunosorbent assay (ELISA). In total, 95 serum samples were analysed, consisting of 30 patients harbouringS. stercoralislarvae, 30 healthy subjects and 35 patients with other parasites. Sensitivity, specificity and diagnostic efficiency were calculated. A significant statistical difference was found in the detection of immune complexes and antibodies in patients harbouringS. stercoralislarvae from larval and eggs antigens, with higher positivity using larval antigen. The larval antigen showed the highest values for sensitivity, specificity and diagnostic efficiency in ELISA from detection of immune complexes. For the first time we used IgG anti-larvae, IgG anti-parasitic females or IgG anti-eggs for immune complex detection. We concluded that the association of antibody and immune complex detection could be used in the diagnosis of human strongyloidiasis.


2014 ◽  
Vol 37 (4) ◽  
pp. 226 ◽  
Author(s):  
Haiyan Liu ◽  
Wenyong Wu ◽  
Yang Liu ◽  
Changle Zhang ◽  
Zheng Zhou

Purpose: Gastric cancer is associated with poor prognosis. The high mortality rate of gastric cancer is mainly attributed to late detection, so diagnosis and treatment are crucial to decreasing mortality. The purpose of this study was to examine the predictive accuracy and discriminative ability of cystic fibrosis transmembrane conductance regulator (CFTR) in gastric cancer patients, in addition to the classical cancer tumor biomarkers carbohydrate antigen 199 (CA199) and carcinoembryonic antigen (CEA). Methods: The study was performed on 78 serum samples from gastric cancer patients and 88 serum samples from healthy adults. Serum levels of CFTR, CA199, CEA and CHN were determined by enzyme-linked immunosorbent assay (ELISA) Results: Spearman’s coefficient analysis showed that, in some cases, CFTR was strongly correlated with CA199 and that CFTR levels increased with age. Kruskal-Wallis testing indicated concentrations of CFTR and CA199 had statistically significant association with stage. Logistic regression showed that CFTR and CA199 independently predicted gastric cancer. Receiver operating characteristics (ROC) showed that combinations of CFTR, CA199, and CEA yielded the best ROC curve, with an AUC of 0.875. Conclusions: The results of this study indicate that the serum CFTR has a broad application prospects for detection of GC.


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