RESUMEN
Sporotrichosis diagnosis involves a series of analyses, including culture and antibody detection in serum samples. Serologic methods may sometimes yield false-negative or false-positive results, leading to inaccurate diagnoses. This study assessed specific patient groups in which antibody detection of different isotypes and subclasses may lack sensitivity. An enzyme-linked immunosorbent assay (ELISA) with Sporothrix brasiliensis exoantigens was used to investigate IgM, IgG, IgG1, IgG2, IgG3, IgG4, IgA, IgA1 and IgA2 antibodies in human serum samples. Eighty serum samples from patients with different sporotrichosis clinical manifestations, including cutaneous forms with and without hypersensitivity manifestations, extracutaneous forms (bone, ocular, meningeal and pulmonary), disseminated cutaneous forms and disseminated forms in individuals living with HIV/AIDS, diabetics and alcoholics, were evaluated. The ELISA sensitivities in the detection of different antibodies ranged from 0.85 to 0.60 for the detection of IgG2 and IgG3, respectively. The antibodies with higher area under ROC curves were IgG2, IgG, IgA and IgA1. There were no significant differences in the immunological reactivity of the tested antibodies among different clinical forms of sporotrichosis. The data revealed a higher likelihood of a false-negative outcome in patients with lesions in the nasal mucosa regarding the detection of IgM and a lower likelihood in patients with lymphocutaneous sporotrichosis regarding the detection of IgG3. Patients with hypersensitivity manifestations had a 3.71 odds ratio to yield negative results in total IgG detection. In conclusion, we identified specific patient groups in which antibody detection may lack sensitivity, thus contributing to a better understanding of the diagnostic challenges associated with this condition.
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Anticuerpos Antifúngicos , Ensayo de Inmunoadsorción Enzimática , Sensibilidad y Especificidad , Sporothrix , Esporotricosis , Humanos , Esporotricosis/inmunología , Esporotricosis/diagnóstico , Anticuerpos Antifúngicos/sangre , Sporothrix/inmunología , Sporothrix/clasificación , Masculino , Femenino , Adulto , Persona de Mediana Edad , Isotipos de Inmunoglobulinas/sangre , Isotipos de Inmunoglobulinas/inmunología , Inmunoglobulina G/sangre , Anciano , Adulto Joven , Antígenos Fúngicos/inmunología , Antígenos Fúngicos/sangre , Inmunoglobulina A/sangre , Inmunoglobulina M/sangreRESUMEN
Antemortem serodiagnosis of aspergillosis remains challenging in Sphenisciformes. Protein electrophoresis, serology (antibody, antigen) by ELISA, and gliotoxin detection provide variable diagnostic value. In the present study, a commercially available Western blot (WB) validated for use in humans and dolphins was adapted for use with penguin samples. Using the same method and reagents, samples were analyzed from multiple institutions in the United States and one facility in France. This was inclusive of normal juvenile African penguins (Spheniscus demersus, n = 10) and various species of penguins in the United States with confirmed infection (n = 9) as well as 52 samples from Humboldt penguins (Spheniscus humboldti) in France. Cumulative WB scores (based on reactivity to different antigens) were found to be significantly higher in the group of penguins with confirmed infection (p < 0.0001). Significant differences were also observed between the clinically normal penguins in the two populations, with higher scores in the United States (median score 1.0, 95%CI [0-5], min 0, max 11) compared to France (median score 0,95%CI [0-0], min 0, max 5). The utilization of the WB as a diagnostic tool is inconclusive due to the use of samples from varying institutions, environmental background, age, and stages of infection. However, this tool may provide an overview of antigen reactivity in penguins infected with Aspergillus to help design a more robust serology assay and further understand the humoral immune response during infection.
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Anticuerpos Antifúngicos , Aspergilosis , Aspergillus , Enfermedades de las Aves , Western Blotting , Spheniscidae , Animales , Aspergilosis/veterinaria , Aspergilosis/diagnóstico , Estados Unidos , Francia , Western Blotting/veterinaria , Aspergillus/inmunología , Anticuerpos Antifúngicos/sangre , Enfermedades de las Aves/diagnóstico , Enfermedades de las Aves/microbiología , Enfermedades de las Aves/inmunologíaRESUMEN
Aspergillus-specific antibodies are diagnostic indicators of allergic bronchopulmonary aspergillosis (ABPA) and chronic pulmonary aspergillosis (CPA). Tests for detecting Aspergillus-specific antibodies were not used clinically in Japan, and the production of the Aspergillus precipitin test was discontinued. Thus, alternative tests for diagnosing aspergillosis are urgently needed. We retrospectively evaluated 64 patients with suspected ABPA and CPA who underwent precipitin antibody testing. Serum Aspergillus IgG levels were measured and compared using the Bordier Aspergillus fumigatus ELISA and the Platelia Aspergillus IgG (Bio-Rad) kits. Of the participants, 18 were diagnosed with CPA, and 8 were diagnosed with ABPA. Both the Bordier and Bio-Rad kits showed high sensitivity and specificity for CPA and ABPA. The area under the receiver operating characteristic curves for the Bordier and Bio-Rad kits were 0.97 and 0.95, respectively, for CPA, and 0.89 and 0.91, respectively, for ABPA. In contrast to the Bordier kit, the Bio-Rad kit showed relatively low anti-Aspergillus IgG levels and lower sensitivity to non-fumigatus Aspergillus infections. The Aspergillus-specific IgG ELISA tests showed sufficient diagnostic accuracy. Therefore, these assays are recommended as alternatives to the precipitin kit for diagnosing aspergillosis in clinical settings in Japan.
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Anticuerpos Antifúngicos , Ensayo de Inmunoadsorción Enzimática , Inmunoglobulina G , Aspergilosis Pulmonar , Sensibilidad y Especificidad , Humanos , Estudios Retrospectivos , Inmunoglobulina G/sangre , Anticuerpos Antifúngicos/sangre , Masculino , Femenino , Persona de Mediana Edad , Anciano , Aspergilosis Pulmonar/diagnóstico , Aspergilosis Pulmonar/inmunología , Adulto , Ensayo de Inmunoadsorción Enzimática/métodos , Japón , Aspergillus/inmunología , Anciano de 80 o más Años , Técnicas para Inmunoenzimas/métodos , Aspergilosis Broncopulmonar Alérgica/diagnóstico , Aspergilosis Broncopulmonar Alérgica/inmunología , Aspergilosis Broncopulmonar Alérgica/sangre , Aspergillus fumigatus/inmunología , Curva ROCRESUMEN
OBJECTIVE: Cryptococcosis predominantly presents as a meningoencephalitis in Thailand. Early and expeditious diagnosis is essential for reducing both mortality and morbidity associated with cryptococcal meningitis. We aim to define and establish the diagnostic performances between the benchmark commercially available diagnostic kit (CrAg® LFA) and the large-scale prototype of an inexpensive in-house immunochromatographic test (ICT) based on monoclonal antibody (MAb) 18B7. METHODS: We have developed the large-scale prototype for the rapid detection of cryptococcal polysaccharide antigens by utilizing a single antibody sandwich ICT format employing MAb 18B7, which is highly specific to Cryptococcus neoformans glucuronoxylomannan (GXM) antigens. An in-house MAb18B7 ICT was manufactured in accordance with industry standards under the control of the International Organization for Standardization (ISO) 13485. RESULTS: The diagnostic sensitivity, specificity, and accuracy for the in-house MAb 18B7 ICT were 99.10%, 97.61%, and 97.83%, respectively. The agreement kappa (κ) coefficient was 0.968 based on the retrospective evaluation of 580 specimens from patients living in northern Thailand with clinically suspected cryptococcosis. CONCLUSION: The data suggest that this in-house MAb 18B7 ICT will be highly beneficial for addressing the issue of cryptococcal infection in Thailand. Moreover, it is anticipated that this inexpensive ICT can play a pivotal role in various global strategies aimed at eradicating cryptococcal meningitis among individuals living with HIV by 2030.
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Anticuerpos Monoclonales , Antígenos Fúngicos , Cromatografía de Afinidad , Criptococosis , Cryptococcus neoformans , Sensibilidad y Especificidad , Humanos , Tailandia , Anticuerpos Monoclonales/inmunología , Cromatografía de Afinidad/métodos , Criptococosis/diagnóstico , Cryptococcus neoformans/inmunología , Cryptococcus neoformans/aislamiento & purificación , Antígenos Fúngicos/análisis , Antígenos Fúngicos/inmunología , Estudios Retrospectivos , Anticuerpos Antifúngicos/sangre , Polisacáridos/análisis , Polisacáridos/inmunología , Masculino , Femenino , Adulto , Pruebas Diagnósticas de Rutina/métodos , Persona de Mediana Edad , Anciano , Adulto JovenRESUMEN
BACKGROUND: Sensitization to Aspergillus fumigatus (AS) has been recently described in chronic obstructive pulmonary disease (COPD) patients. However, there is no data on the community prevalence of AS in COPD. OBJECTIVES: To assess the prevalence of AS among COPD subjects. The secondary objectives were to (1) assess the prevalence of allergic bronchopulmonary aspergillosis (ABPA) in COPD and (2) compare the lung function in COPD subjects with and without AS. METHODS: We conducted a cross-sectional study in rural (29 villages) and urban (20 wards) communities in North India. We identified individuals with respiratory symptoms (IRS) through a house-to-house survey using a modified IUATLD questionnaire. We then diagnosed COPD through specialist assessment and spirometry using the GOLD criteria. We assayed A.fumigatus-specific IgE in COPD subjects. In those with A. fumigatus-specific IgE ≥0.35 kUA/L (AS), ABPA was diagnosed with raised serum total IgE and raised A.fumigatus-specific IgG or blood eosinophil count. RESULTS: We found 1315 (8.2%) IRS among 16,071 participants >40 years and diagnosed COPD in 355 (2.2%) subjects. 291 (82.0%) were men and 259 (73.0%) resided in rural areas. The prevalence of AS and ABPA was 17.7% (95% CI, 13.9-21.8) and 6.6% (95% CI, 4.4-8.8). We found a lower percentage predicted FEV1 in COPD subjects with AS than those without (p =.042). CONCLUSIONS: We found an 18% community prevalence of AS in COPD subjects in a specific area in North India. Studies from different geographical areas are required to confirm our findings. The impact of AS and ABPA on COPD requires further research.
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Aspergilosis Broncopulmonar Alérgica , Aspergillus fumigatus , Inmunoglobulina E , Enfermedad Pulmonar Obstructiva Crónica , Humanos , Enfermedad Pulmonar Obstructiva Crónica/complicaciones , Enfermedad Pulmonar Obstructiva Crónica/epidemiología , India/epidemiología , Masculino , Estudios Transversales , Femenino , Aspergilosis Broncopulmonar Alérgica/epidemiología , Persona de Mediana Edad , Prevalencia , Aspergillus fumigatus/inmunología , Anciano , Adulto , Inmunoglobulina E/sangre , Anticuerpos Antifúngicos/sangre , Población Rural/estadística & datos numéricos , Población Urbana/estadística & datos numéricosRESUMEN
PURPOSE: This study aimed to develop a double antigen sandwich ELISA (DAgS-ELISA) method for more efficient, accurate, and quantitative detection of total antibodies against Candida albicans enolase1 (CaEno1) for diagnosing invasive candidiasis (IC). METHODS: DAgS-ELISA was developed using recombinant CaEno1 and a monoclonal antibody as the standard. Performance evaluation included limit of detection, accuracy, and repeatability. Dynamic changes in antibody levels against CaEno1 in serum from systemic candidiasis mice were analyzed using DAgS-ELISA. Patient serum samples from IC, Candida colonization, bacterial infections, and healthy controls were analyzed with DAgS-ELISA and indirect ELISA. RESULTS: DAgS-ELISA outperformed indirect ELISA in terms of linear range and test background. In systemic candidiasis mice, a distinctive 'double-peak' pattern in dynamic antibody levels was observed. Additionally, there was a high level of consistency in the positive rates of CaEno1 antibodies detected by both DAgS-ELISA and indirect ELISA. While the positivity rates differed among patient groups, no significant variations in antibody levels were detected among the various positive patient groups. CONCLUSIONS: DAgS-ELISA offers a reliable novel approach for IC diagnosis, enabling rapid, accurate, and quantitative detection of CaEno1 antibodies. Further validation and optimization are needed for its clinical application and effectiveness.
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Anticuerpos Antifúngicos , Candida albicans , Ensayo de Inmunoadsorción Enzimática , Fosfopiruvato Hidratasa , Ensayo de Inmunoadsorción Enzimática/métodos , Animales , Fosfopiruvato Hidratasa/inmunología , Fosfopiruvato Hidratasa/sangre , Candida albicans/inmunología , Anticuerpos Antifúngicos/sangre , Ratones , Humanos , Candidiasis Invasiva/diagnóstico , Candidiasis Invasiva/inmunología , Candidiasis Invasiva/sangre , Femenino , Candidiasis/diagnóstico , Candidiasis/sangre , Candidiasis/inmunología , Antígenos Fúngicos/inmunología , Antígenos Fúngicos/sangre , Sensibilidad y Especificidad , Proteínas Fúngicas/inmunología , Anticuerpos Monoclonales/inmunología , Ratones Endogámicos BALB CRESUMEN
CONTEXT: Allergic bronchopulmonary mycoses (ABPM) can be due to molds other than Aspergillus fumigatus in patients with cystic fibrosis (pwCF). We aimed to develop immunoassays for the detection of specific IgE (sIgE) directed against five fungal species involved in ABPM: Aspergillus terreus, Scedosporium apiospermum, Lomentospora prolificans, Rasamsonia argillacea, and Exophiala dermatitidis. MATERIALS AND METHODS: Serum samples (n = 356) from 238 pwCF, collected in eight CF care centers in France, Germany, and Italy, were analyzed by dissociated enhanced lanthanide fluorescent immunoassay (DELFIA®) to assess levels of sIgE directed against antigenic extracts of each fungus. Clinical, biological, and radiological data were collected for each episode. One hundred serum samples from healthy blood donors were used as controls. Sera were classified into four groups depending on the level of sIgE according to the quartile repartition calculated for the pwCF population. A score of 4 for values above the 3rd quartile corresponds to an elevated level of sIgE. RESULTS: PwCF showed higher levels of sIgE than controls. Based on criteria from the ABPA-ISHAM working group, with an additional criterion of "a sIgE score of 4 for at least one non-A. fumigatus mold", we were able to diagnose six cases of ABPM. CONCLUSIONS: Using 417 IU/mL as the threshold for total IgE and the same additional criterion, we identified seven additional pwCF with "putative ABPM". Detection of sIgE by DELFIA® showed good analytical performance and supports the role played by non-A. fumigatus molds in ABPM. However, commercially available kits usable in routine practice are needed to improve the diagnosis of ABPM.
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Anticuerpos Antifúngicos , Fibrosis Quística , Hongos , Inmunoglobulina E , Humanos , Fibrosis Quística/complicaciones , Inmunoglobulina E/sangre , Femenino , Masculino , Adulto , Adulto Joven , Adolescente , Hongos/inmunología , Hongos/clasificación , Hongos/aislamiento & purificación , Inmunoensayo/métodos , Niño , Anticuerpos Antifúngicos/sangre , Italia , Francia , Alemania , Preescolar , Persona de Mediana Edad , Aspergilosis Broncopulmonar Alérgica/diagnóstico , Aspergilosis Broncopulmonar Alérgica/inmunología , Aspergilosis Broncopulmonar Alérgica/sangreRESUMEN
Inflammatory bowel disease (IBD) is characterized by chronic inflammation in the gut. There is growing evidence in Crohn's disease (CD) of the existence of a preclinical period characterized by immunological changes preceding symptom onset that starts years before diagnosis. Gaining insight into this preclinical phase will allow disease prediction and prevention. Analysis of preclinical serum samples, up to 6 years before IBD diagnosis (from the PREDICTS cohort), revealed the identification of a unique glycosylation signature on circulating antibodies (IgGs) characterized by lower galactosylation levels of the IgG fragment crystallizable (Fc) domain that remained stable until disease diagnosis. This specific IgG2 Fc glycan trait correlated with increased levels of antimicrobial antibodies, specifically anti-Saccharomyces cerevisiae (ASCA), pinpointing a glycome-ASCA hub detected in serum that predates by years the development of CD. Mechanistically, we demonstrated that this agalactosylated glycoform of ASCA IgG, detected in the preclinical phase, elicits a proinflammatory immune pathway through the activation and reprogramming of innate immune cells, such as dendritic cells and natural killer cells, via an FcγR-dependent mechanism, triggering NF-κB and CARD9 signaling and leading to inflammasome activation. This proinflammatory role of ASCA was demonstrated to be dependent on mannose glycan recognition and galactosylation levels in the IgG Fc domain. The pathogenic properties of (anti-mannose) ASCA IgG were validated in vivo. Adoptive transfer of antibodies to mannan (ASCA) to recipient wild-type mice resulted in increased susceptibility to intestinal inflammation that was recovered in recipient FcγR-deficient mice. Here we identify a glycosylation signature in circulating IgGs that precedes CD onset and pinpoint a specific glycome-ASCA pathway as a central player in the initiation of inflammation many years before CD diagnosis. This pathogenic glyco-hub may constitute a promising new serum biomarker for CD prediction and a potential target for disease prevention.
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Enfermedad de Crohn , Inmunoglobulina G , Manosa , Polisacáridos , Enfermedad de Crohn/inmunología , Enfermedad de Crohn/sangre , Inmunoglobulina G/inmunología , Inmunoglobulina G/sangre , Animales , Humanos , Glicosilación , Manosa/metabolismo , Manosa/inmunología , Ratones , Polisacáridos/inmunología , Polisacáridos/metabolismo , Femenino , Saccharomyces cerevisiae/inmunología , Masculino , Adulto , Anticuerpos Antifúngicos/sangre , Anticuerpos Antifúngicos/inmunología , Ratones Endogámicos C57BL , Ratones Noqueados , Biomarcadores/sangre , Persona de Mediana Edad , Fragmentos Fc de Inmunoglobulinas/inmunología , GlicoproteínasRESUMEN
Introduction: Equine asthma (EA) is a common lower airway disease in horses, but whether its pathogenesis is allergic is ambiguous. Extrinsic stimuli like hay dust induce acute exacerbation of clinical signs and sustained local neutrophilic inflammation in susceptible horses. Aspergillus fumigatus is an EA stimulus, but it is unclear if it merely acts as an IgE-provoking allergen. We aimed to comprehensively analyze immunoglobulin (Ig) isotypes in EA, elucidating their binding to different A. fumigatus antigens, and their quantities systemically in serum and locally in bronchoalveolar lavage fluid (BALF). Methods: Serum and BALF from healthy horses (HE, n = 18) and horses with mild-moderate asthma (MEA, n = 20) or severe asthma (SEA, n = 24) were compared. Ig isotype (IgG1, IgG3/5, IgG4/7, IgG6, IgA, and IgE) binding to nine antigens (A. fumigatus lysate, and recombinant Asp f 1, Asp f 7, Asp f 8, dipeptidyl-peptidase 5, class II aldolase/adducin domain protein, glucoamylase, beta-hexosaminidase, and peptide hydrolase) was compared by enzyme-linked immunosorbent assays. Total Ig isotype contents were determined by bead-based assays. Results: MEA and SEA differed from HE but hardly from each other. Compared to HE, asthmatic horses showed increased anti-A. fumigatus binding of IgG (BALF and serum) and IgA (BALF). Serum and BALF IgE binding and total IgE contents were similar between HE and EA. Single antigens, as well as A. fumigatus lysate, yielded similar Ig binding patterns. Serum and BALF IgG1 binding to all antigens was increased in SEA and to several antigens in MEA. Serum IgG4/7 binding to two antigens was increased in SEA. BALF IgA binding to all antigens was increased in SEA and MEA. Total BALF IgG1 and IgG4/7 contents were increased in SEA, and serum IgG4/7 content was increased in MEA compared to HE. Yet, total isotype contents differentiated EA and HE less clearly than antigen-binding Ig. Discussion: A. fumigatus immunogenicity was confirmed without identification of single dominant antigens here. A. fumigatus provoked elevated BALF IgG1 and IgA binding, and these isotypes appear relevant for neutrophilic EA, which does not support allergy. BALF Ig isotype differentiation beyond IgE is crucial for a comprehensive analysis of immune responses to fungi in EA pathogenesis.
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Antígenos Fúngicos , Aspergillus fumigatus , Asma , Líquido del Lavado Bronquioalveolar , Enfermedades de los Caballos , Inmunoglobulina A , Inmunoglobulina G , Animales , Caballos/inmunología , Aspergillus fumigatus/inmunología , Líquido del Lavado Bronquioalveolar/inmunología , Asma/inmunología , Asma/microbiología , Inmunoglobulina G/inmunología , Inmunoglobulina G/sangre , Inmunoglobulina A/inmunología , Inmunoglobulina A/sangre , Inmunoglobulina A/metabolismo , Enfermedades de los Caballos/inmunología , Enfermedades de los Caballos/microbiología , Antígenos Fúngicos/inmunología , Masculino , Neutrófilos/inmunología , Neutrófilos/metabolismo , Femenino , Inmunoglobulina E/inmunología , Inmunoglobulina E/sangre , Anticuerpos Antifúngicos/inmunología , Anticuerpos Antifúngicos/sangreRESUMEN
OBJECTIVES: The aim of the present study was to evaluate minimally invasive diagnostic techniques, such as the semi-quantitative indirect IgG antibody enzyme immunoassay (EIA) using blood serum and the urinary lateral flow assay (LFA), for the detection of Histoplasma capsulatum in cats with histoplasmosis. METHODS: Eight client-owned domestic cats diagnosed with histoplasmosis were selected based on cytological, histopathological, mycological, molecular or antigenic techniques. The blood serum of these animals was tested in a semi-quantitative indirect IgG antibody EIA for the detection of H capsulatum. Urine samples were tested for H capsulatum antigen using LFA. RESULTS: Five cats were seropositive on IgG EIA (5/8, with diagnostic sensitivity equal to 62.5%; 95% confidence interval [CI] 24.5-91.5) and five cats were positive on H capsulatum antigen LFA (5/7, with diagnostic sensitivity equal to 71.4%; 95% CI 29.0-96.3). The combined diagnostic sensitivity when interpreted in parallel was 87.5% (7/8, 95% CI 47.3-99.7). The specificity for the anti-Histoplasma IgG EIA was 100% (95% CI 71.5-100) and for the H capsulatum antigen LFA it was also 100% (95% CI 71.5-100). CONCLUSIONS AND RELEVANCE: The semi-quantitative indirect IgG antibody EIA for the detection of H capsulatum in blood serum and the urinary LFA for the detection of the same agent emerge as new minimally invasive diagnostic techniques that can assist in the approach to disseminated and pulmonary feline histoplasmosis, especially when both techniques are considered together.
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Enfermedades de los Gatos , Histoplasma , Histoplasmosis , Sensibilidad y Especificidad , Gatos , Animales , Histoplasmosis/veterinaria , Histoplasmosis/diagnóstico , Enfermedades de los Gatos/diagnóstico , Enfermedades de los Gatos/microbiología , Histoplasma/aislamiento & purificación , Histoplasma/inmunología , Masculino , Femenino , Anticuerpos Antifúngicos/sangre , Técnicas para Inmunoenzimas/veterinaria , Inmunoglobulina G/sangreRESUMEN
In the present study, we validate and compare the second-generation Miravista Coccidioides IgG and IgM enzyme immunoassays (EIA) (MiraVista Diagnostics [MVD] Ab EIA) to Meridian Diagnostics Coccidioides IgG and IgM EIA (Meridian Ab EIA), immunodiffusion (ID) and complement fixation (CF). We also evaluated whether the addition of Coccidioides antigen testing to anti-Coccidioides antibody testing increased the sensitivity for the diagnosis of currently active coccidioidomycosis. We retrospectively studied 555 patients evaluated at Valleywise Health Medical Center between January 2013 and May 2017 for whom coccidioidomycosis was suspected and samples were submitted to MVD for testing. Specimens were tested for antigen in the MVD antigen enzyme immunoassay (MVD Ag EIA) and for IgG and IgM antibodies with MVD and Meridian Diagnostics EIAs. ID and CF were obtained from medical records. Sensitivity and specificity were 83.0% and 91.1% or MVD Ab EIA, 69.3% and 99.7% for Meridian Ab EIA, 85.4% and 100% for ID and 65.5% and 100% for CF. Combined MVD antigen and antibody detection by EIA and ID resulted in increased sensitivity in disseminated and pulmonary disease (MVD Ag/MVD Ab: 100%, 88.3%; MVD Ag/Meridian Ab: 98.2%, 78.6%; and MVD Ag/ID: 100%, 91.7%). The detection of antibodies by MVD EIA was more sensitive than Meridian EIA or CF but similar to ID. This study supports the use of antigen testing in immunocompromised patients and those with suspected disseminated disease. Furthermore, the addition of antigen detection by EIA to antibody detection resulted in higher sensitivity of all serological tests.
The most common methods for the diagnosis of moderate or severe coccidioidomycosis rely on the detection of antibodies or antigens. Here we present the validation of a new Miravista Coccidioides antibody detection test combined with antigen detection and compare it to other immunodiagnostics.
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Anticuerpos Antifúngicos , Antígenos Fúngicos , Coccidioides , Coccidioidomicosis , Técnicas para Inmunoenzimas , Inmunoglobulina G , Inmunoglobulina M , Sensibilidad y Especificidad , Humanos , Coccidioidomicosis/diagnóstico , Coccidioidomicosis/inmunología , Antígenos Fúngicos/inmunología , Antígenos Fúngicos/sangre , Inmunoglobulina M/sangre , Estudios Retrospectivos , Inmunoglobulina G/sangre , Coccidioides/inmunología , Técnicas para Inmunoenzimas/métodos , Anticuerpos Antifúngicos/sangre , Masculino , Persona de Mediana Edad , Femenino , Anciano , Adulto , Adolescente , Adulto Joven , Niño , Anciano de 80 o más Años , Preescolar , Inmunodifusión , LactanteAsunto(s)
Aspergilosis , Aspergillus , Análisis Costo-Beneficio , Inmunoglobulina G , Inmunoglobulina M , Humanos , Uganda , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Aspergilosis/diagnóstico , Anticuerpos Antifúngicos/sangre , Pruebas Serológicas/métodos , Pruebas Serológicas/economía , Análisis de Costo-EfectividadRESUMEN
BACKGROUND: Aspergillus is one of the most common pathogens causing fungal allergy in the respiratory tract. Serum Aspergillus fumigatus-specific immunoglobulin G (Af-sIgG) levels have been used as a biomarker for the diagnosis and treatment response monitoring in airway allergic diseases such as allergic bronchopulmonary aspergillosis and allergic fungal rhinosinusitis. However, its role in common primary chronic rhinosinusitis (CRS) was unclear. OBJECTIVE: This study aims to evaluate whether serum Af-sIgG level could serve as a biomarker for the disease presentation of primary CRS. METHODS: We obtained serum Af-sIgG levels from patients diagnosed as bilateral primary CRS refractory to medical treatment and evaluated the correlations between serum Af-sIgG levels and disease severity in patients with type 2 (T2) and non-T2 CRS. RESULTS: Patients with T2 CRS exhibited significantly higher serum Af-sIgG levels than non-T2 CRS patients. The cut-off value of serum Af-sIgG in T2 CRS was 20.9â mg/L, with an odds ratio of 3.8 (95% CI 1.17-12.20, P = .026). Furthermore, serum Af-sIgG levels were positively correlated with symptom scores evaluated by the Sino-Nasal Outcome Test-22 (SNOT-22) scores in T2 patients (P = .009). While stratified by SNOT-22 total scores, patients with severe disease had higher serum Af-sIgG levels only in T2 CRS (P = .034). In individual domains of SNOT-22 analysis, serum Af-sIgG levels showed a significant correlation with "ear/facial" symptom scores in the T2 group (P < .001). CONCLUSIONS: Serum Af-sIgG levels may serve as a supplementary objective biomarker that correlates with identification and subjective measurements of T2 CRS, and may be associated with symptoms arising from Eustachian tube dysfunction.
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Anticuerpos Antifúngicos , Aspergillus fumigatus , Biomarcadores , Inmunoglobulina G , Rinitis , Sinusitis , Humanos , Sinusitis/diagnóstico , Sinusitis/inmunología , Sinusitis/sangre , Sinusitis/microbiología , Inmunoglobulina G/sangre , Aspergillus fumigatus/inmunología , Biomarcadores/sangre , Enfermedad Crónica , Rinitis/diagnóstico , Rinitis/inmunología , Rinitis/sangre , Masculino , Femenino , Persona de Mediana Edad , Adulto , Anticuerpos Antifúngicos/sangre , Anciano , Aspergilosis/diagnóstico , Aspergilosis/inmunología , Aspergilosis/sangre , Índice de Severidad de la Enfermedad , RinosinusitisRESUMEN
BACKGROUND: Chronic pulmonary aspergillosis (CPA) is known to complicate patients with post-tubercular lung disease. However, some evidence suggests that CPA might co-exist in patients with newly-diagnosed pulmonary tuberculosis (P.TB) at diagnosis and also develop during therapy. The objective of this study was to confirm the presence of CPA in newly diagnosed P.TB at baseline and at the end-of-TB-therapy. MATERIALS AND METHODS: This prospective longitudinal study included newly diagnosed P.TB patients, followed up at third month and end-of-TB-therapy with symptom assessment, anti-Aspergillus IgG antibody and imaging of chest for diagnosing CPA. RESULTS: We recruited 255 patients at baseline out of which 158 (62%) completed their follow-up. Anti-Aspergillus IgG was positive in 11.1% at baseline and 27.8% at end-of-TB-therapy. Overall, proven CPA was diagnosed in 7% at baseline and 14.5% at the end-of-TB-therapy. Around 6% patients had evidence of aspergilloma in CT chest at the end-of-TB-therapy. CONCLUSIONS: CPA can be present in newly diagnosed P.TB patients at diagnosis and also develop during anti-tubercular treatment. Patients with persistent symptoms or developing new symptoms during treatment for P.TB should be evaluated for CPA. Whether patients with concomitant P.TB and CPA, while receiving antitubercular therapy, need additional antifungal therapy, needs to be evaluated in future studies.
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Aspergilosis Pulmonar , Tuberculosis Pulmonar , Humanos , Masculino , Femenino , Aspergilosis Pulmonar/epidemiología , Aspergilosis Pulmonar/tratamiento farmacológico , Aspergilosis Pulmonar/complicaciones , Aspergilosis Pulmonar/diagnóstico , Tuberculosis Pulmonar/tratamiento farmacológico , Tuberculosis Pulmonar/complicaciones , Tuberculosis Pulmonar/epidemiología , Tuberculosis Pulmonar/diagnóstico , Persona de Mediana Edad , Estudios Prospectivos , Adulto , Estudios Longitudinales , Incidencia , Anciano , Anticuerpos Antifúngicos/sangre , Enfermedad Crónica , Estudios de Seguimiento , Inmunoglobulina G/sangre , Antituberculosos/uso terapéutico , Aspergillus/aislamiento & purificación , Aspergillus/inmunología , Adulto JovenRESUMEN
BACKGROUND: Accurate diagnosis of paracoccidioidomycosis is crucial for improving patient outcomes. Paracoccidioides antibody detection by double immunodiffusion (DID) is a convenient diagnostic tool, but testing performance can vary based on certain factors. METHODS: We assessed DID performance using a commercially prepared Paracoccidioides reagents (IMMY, USA), involving 40 serum specimens, including 20 from patients with proven paracoccidioidomycosis and 20 from patients without the disease. The DID test demonstrated a sensitivity of 90% (95% CI=68%-99%) and a specificity of 100% (95% CI=83%-100%). CONCLUSIONS: Our findings suggest that DID using commercial reagents may provide a feasible tool with satisfactory testing performance for anti-Paracoccidioides antibody detection.
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Anticuerpos Antifúngicos , Inmunodifusión , Paracoccidioides , Paracoccidioidomicosis , Sensibilidad y Especificidad , Humanos , Anticuerpos Antifúngicos/sangre , Paracoccidioidomicosis/diagnóstico , Paracoccidioidomicosis/inmunología , Paracoccidioides/inmunología , Juego de Reactivos para Diagnóstico , Femenino , MasculinoRESUMEN
Brazil-grown outdoor-cultivated Agaricus brasiliensis KA21 fruiting body (KA21) significantly increases the production of serum anti-beta-glucan antibody. Therefore, KA21 ingestion may be useful for the prevention and alleviation of fungal infections. This study aimed to determine the effects of KA21 in fungal infections in animals. KA21 was administered to nine dogs infected with Malassezia. Notably, the anti-beta-glucan antibody titer remained unchanged or tended to decrease in the oral steroid arm, whereas in the non-steroid arm, antibody titer increased in almost all animals after KA21 ingestion. Dogs showing improved clinical symptoms exhibited increased anti-beta-glucan antibody titers. The results of this study suggest that KA21 ingestion may alleviate the symptoms of Malassezia and other fungal infections and that continuous ingestion may help prolong recurrence-free intervals. Additionally, the ingestion of KA21 during oral steroid dosage reduction or discontinuation may enable smoother steroid withdrawal.
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Agaricus , Enfermedades de los Perros , Cuerpos Fructíferos de los Hongos , Malassezia , Animales , Perros , Agaricus/química , Cuerpos Fructíferos de los Hongos/química , Malassezia/efectos de los fármacos , Enfermedades de los Perros/microbiología , Enfermedades de los Perros/tratamiento farmacológico , Dermatomicosis/veterinaria , Dermatomicosis/prevención & control , Dermatomicosis/tratamiento farmacológico , Dermatomicosis/microbiología , beta-Glucanos/administración & dosificación , beta-Glucanos/farmacología , Masculino , Brasil , Dermatitis/tratamiento farmacológico , Dermatitis/veterinaria , Dermatitis/microbiología , Dermatitis/prevención & control , Femenino , Anticuerpos Antifúngicos/sangreRESUMEN
BACKGROUND: Due to a delay in diagnosis by conventional techniques and high mortality, the development of a standardised and rapid non-culture-based technique is an unmet need in pulmonary, gastrointestinal, and disseminated forms of mucormycosis. Though limited studies have been conducted for molecular diagnosis, there are no established serologic tests for this highly fatal infection. OBJECTIVE: To develop and evaluate an indirect in-house enzyme-linked immunosorbent assay (ELISA) utilising antigens of Rhizopus arrhizus for detecting anti-Rhizopus antibodies (IgG and IgM) in sera of patients with mucormycosis. METHODS: We extracted both secretory and mycelial Rhizopus antigens using standardised protocols. Bradford assay was used for protein quantification. We then standardised an indirect ELISA using R. arrhizus mycelial and secretory antigens (10.0 µg/mL in bicarbonate buffer pH 9.2) for detecting anti-Rhizopus IgG and IgM antibodies in patient sera. We included patients with mucormycosis, other fungal infections, and healthy controls. Antibody index value (E-value) was calculated for each patient sample. RESULTS: Asparagine broth culture filtrate utilising 85% ammonium sulphate salt fractionation and mycelial homogenate grown in yeast extract peptone dextrose (YPD) broth precipitated with trichloroacetic acid (TCA) yielded a large amount of good-quality protein for the assay. We included 55 patients with mucormycosis (rhino-orbito-cerebral mucormycosis [ROCM, n = 39], pulmonary [n = 15], gastrointestinal [n = 1]), 24 with other fungal infections (probable aspergillosis [n = 14], candidiasis [n = 10]), and healthy controls (n = 16). The sensitivity of the antibody test for diagnosing mucormycosis ranged from 83.6-92.7% for IgG and 72.7-87.3% for IgM, with a specificity of 91.7-92.5% for IgG and 80-82.5% for IgM. The sera from patients with other fungal infections and healthy individuals did not show significant cross-reactivity. CONCLUSION: The detection of anti-Rhizopus IgG antibody performed significantly better in comparison to IgM-based ELISA for diagnosing both ROCM (sensitivity of 84.6% vs. 69.2%) and pulmonary cases (86.6% vs. 80.0%). More extensive studies are required to confirm our findings.
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Anticuerpos Antifúngicos , Antígenos Fúngicos , Ensayo de Inmunoadsorción Enzimática , Inmunoglobulina G , Inmunoglobulina M , Mucormicosis , Rhizopus , Sensibilidad y Especificidad , Pruebas Serológicas , Mucormicosis/diagnóstico , Mucormicosis/microbiología , Mucormicosis/inmunología , Humanos , Rhizopus/inmunología , Ensayo de Inmunoadsorción Enzimática/métodos , Antígenos Fúngicos/inmunología , Antígenos Fúngicos/análisis , Pruebas Serológicas/métodos , Anticuerpos Antifúngicos/sangre , Inmunoglobulina M/sangre , Inmunoglobulina G/sangre , Femenino , Masculino , Persona de Mediana EdadRESUMEN
The delay in making a correct diagnosis of Candida auris causes concern in the healthcare system setting, and immunoproteomics studies are important to identify immunoreactive proteins for new diagnostic strategies. In this study, immunocompetent murine systemic infections caused by non-aggregative and aggregative phenotypes of C. auris and by Candida albicans and Candida haemulonii were carried out, and the obtained sera were used to study their immunoreactivity against C. auris proteins. The results showed higher virulence, in terms of infection signs, weight loss, and histopathological damage, of the non-aggregative isolate. Moreover, C. auris was less virulent than C. albicans but more than C. haemulonii. Regarding the immunoproteomics study, 13 spots recognized by sera from mice infected with both C. auris phenotypes and analyzed by mass spectrometry corresponded to enolase, phosphoglycerate kinase, glyceraldehyde-3-phosphate dehydrogenase, and phosphoglycerate mutase. These four proteins were also recognized by sera obtained from human patients with disseminated C. auris infection but not by sera obtained from mice infected with C. albicans or Aspergillus fumigatus. Spot identification data are available via ProteomeXchange with the identifier PXD049077. In conclusion, this study showed that the identified proteins could be potential candidates to be studied as new diagnostic or even therapeutic targets for C. auris.
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Candida , Candidiasis , Inmunoglobulina G , Animales , Ratones , Candida/inmunología , Candida/patogenicidad , Humanos , Candidiasis/inmunología , Candidiasis/microbiología , Candidiasis/sangre , Inmunoglobulina G/sangre , Antígenos Fúngicos/inmunología , Antígenos Fúngicos/sangre , Proteómica/métodos , Candida albicans/inmunología , Candida albicans/patogenicidad , Proteínas Fúngicas/inmunología , Fosfoglicerato Mutasa/inmunología , Fosfoglicerato Quinasa/inmunología , Gliceraldehído-3-Fosfato Deshidrogenasas/inmunología , Gliceraldehído-3-Fosfato Deshidrogenasas/metabolismo , Anticuerpos Antifúngicos/sangre , Anticuerpos Antifúngicos/inmunología , Femenino , VirulenciaRESUMEN
The detection of patterns associated with the invasive form of Candida albicans, such as Candida albicans germ tube antibodies (CAGTA), is a useful complement to blood culture for Invasive Candidiasis (IC) diagnosis. As CAGTA are detected by a non-standardisable and non-automatable technique, a Candida albicans cDNA expression library was screened with CAGTA isolated from serum of an animal model of invasive candidiasis, and five protein targets were identified: hyphally regulated cell wall protein 1 (Hyr1), enolase 1 (Eno1), coatomer subunit gamma (Sec21), a metallo-aminopeptidase (Ape2) and cystathionine gamma-lyase (Cys3). Homology with proteins from other organisms rules out Cys3 as a good biomarker while Sec21 results suggest that it is not in the germ tubes surface but secreted to the external environment. Our analysis propose Ape2, Sec21 and a region of Hyr1 different from the one currently being studied for immunoprotection as potential biomarker candidates for the diagnosis of IC.
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Anticuerpos Antifúngicos , Candida albicans , Candidiasis Invasiva , Proteínas Fúngicas , Biblioteca de Genes , Candida albicans/genética , Candidiasis Invasiva/diagnóstico , Candidiasis Invasiva/microbiología , Animales , Proteínas Fúngicas/genética , Anticuerpos Antifúngicos/sangre , Biomarcadores/sangre , Modelos Animales de Enfermedad , Humanos , RatonesRESUMEN
INTRODUCTION: Aspergillus fumigatus is the most common airborne allergen of the Aspergillus family. However, allergies to Aspergillus spp. are increasing, and subsequently, allergies to Aspergillus species other than fumigatus are also on the rise. Commercial diagnostic tools are still limited to Aspergillus fumigatus. Hence, there is a need for improved tests. We decided to investigate the correlation between serological sensitization to A. fumigatus and other Aspergillus species. METHODS: Hundred and seven patients with positive skin prick tests to A. fumigatus were included in this study. Immunoglobulin E (IgE) concentrations against A. fumigatus, A. terreus, A. niger, A. flavus, and A. versicolor were measured from specimens by fluorescent enzyme-linked immunoassays. RESULTS: Patients showed considerably higher IgE concentrations against A. fumigatus (6.00 ± 15.05 kUA/L) than A. versicolor (0.30 ± 1.01 kUA/L), A. niger (0.62 ± 1.59 kUA/L), A. terreus (0.45 ± 1.12 kUA/L), or A. flavus (0.41 ± 0.97 kUA/L). Regression analysis yielded weak positive correlations for all Aspergillus spp., but low r2 values and heteroscedastic distribution indicate an overall poor fit of the calculated models. CONCLUSION: Serological sensitization against A. fumigatus does not correlate with sensitization against other Aspergillus spp. To detect sensitization against these, other diagnostic tools like a skin prick test solution of different Aspergillus spp. are needed.