Adult ; Alternaria ; immunology ; Alveolitis, Extrinsic Allergic ; immunology ; Animals ; Antibodies ; immunology ; Antibodies, Bacterial ; immunology ; Antibodies, Fungal ; immunology ; Antigens ; immunology ; Antigens, Bacterial ; immunology ; Antigens, Fungal ; immunology ; Aspergillus fumigatus ; immunology ; Asymptomatic Diseases ; Candida albicans ; immunology ; Cladosporium ; immunology ; Columbidae ; immunology ; Female ; Healthy Volunteers ; Humans ; Immunoglobulin G ; immunology ; Male ; Melopsittacus ; immunology ; Middle Aged ; Mucor ; immunology ; Nocardia ; immunology ; Parrots ; immunology ; Penicillium chrysogenum ; immunology ; Stachybotrys ; immunology ; Thermoactinomyces ; immunology

OBJECTIVETo establish an immunological method for detecting antibodies of Penicillium marneffei.

METHODSThe recombinant Mp1p protein of Penicillium marneffei was expressed in Pichia pastoris and labeled with HRP (Mp1p-HRP) with a modified sodium periodate method. A double-antigen sandwich enzyme-linked immunosorbant assay (ELISA) was established by determining the optimal coating concentration of Mp1p protein and the concentration of the detecting protein Mp1p-HRP. The sensitivity and specificity of the assay was evaluated by detecting Mp1p antibodies in 100 serum samples from healthy donors, 15 samples from culture-confirmed penicilliosis patients, and 21 samples from patients with culture-confirmed other fungal infections.

RESULTSA double-antigen sandwich ELISA was successfully established for detecting Mp1p-specific antibody. The specificity of the assay was 100% (121/121) for detecting Mp1p-specific antibody in the sera from healthy donors and patients with other fungal infection. The detection results of the 15 serum samples from patients with culture-confirmed penicilliosis showed positivity for Mp1p antibody in 2 samples and Mp1p antigen positivity in 12 samples; combining the detection results of Mp1p antigen and antibody obviously increased the diagnostic sensitivity to 93.3% (14/15).

CONCLUSIONThe double-antigen sandwich ELISA shows a high specificity in detecting Mp1p-specific antibody, and simultaneous detection of Mp1p antigen and antibody can increase the diagnostic sensitivity for penicilliosis.

Antibodies, Fungal ; blood ; immunology ; Antigens, Fungal ; blood ; immunology ; Enzyme-Linked Immunosorbent Assay ; methods ; Humans ; Mycoses ; blood ; diagnosis ; microbiology ; Penicillium ; immunology ; Pichia ; immunology ; Sensitivity and Specificity

We report a case of hypersensitivity pneumonitis in a 30-yr-old female housewife caused by Penicillium species found in her home environment. The patient was diagnosed according to history, chest radiograph, spirometry, high-resolution chest CT, and transbronchial lung biopsy. To identify the causative agent, cultured aeromolds were collected by the open-plate method. From the main fungi cultured, fungal antigens were prepared, and immunoblot analysis with the patient's serum and each fungal antigen was performed. A fungal colonies were isolated from the patient's home. Immunoblotting analysis with the patient's sera demonstrated a IgG-binding fractions to Penicillium species extract, while binding was not noted with control subject. This study indicates that the patient had hypersensitivity pneumonitis on exposure to Penicillium species in her home environment.
Adult ; Alveolitis, Extrinsic Allergic/*etiology/immunology/*microbiology ; Antibodies, Fungal/blood ; Antigens, Fungal ; Environmental Microbiology ; Female ; Housing ; Humans ; Immunoglobulin G/blood ; Korea ; Penicillium/*immunology/isolation and purification/*pathogenicity

OBJECTIVETo evaluate the diagnostic value of serum galactomannan antigen (GM) and (1→3)-β-D-glucan antigen (BG) assay in invasive fungal infections (IFI) in the patients with hematologic malignancies and the role in monitoring therapeutic response.

METHODSFifty one patients with hematological malignancies met the criteria for inclusion: (1) body temperature above 38°C for 48 hours, (2) failure to respond to broad-spectrum antibiotic treatment, or (3) temperature rose again after the responded drop. Blood samples were collected twice at the first week, then once a week in at least four weeks. The double antibody sandwich enzyme-linked immunosorbent assay (ELISA) and colorimetric assay were used for detecting GM and BG. The positive GM test is defined as two consecutive tests at different time GM value > 0.5 or > 0.8 and the positive G test is defined as BG value > 80 pg/ml. The patients were assigned into four groups as proven, probable, possible, and non-fungal infection respectively, and 21 normal volunteers were as controls.

RESULTSTwo hundred and forty serum samples were collected from 51 patients including 2 of proven IFI, 26 probable IFI, 17 possible IFI and 6 non-fungal infection. The true-positive group including the proven and probable groups, and true negative group was the non-fungal infection group. GM tests were positive in 21 of 28 cases in true positive group, and only one of 6 cases in non-fungal infection. The sensitivity, specificity, positive predictive value and negative predictive value were 75%, 83.3%, 95.5% and 41.7%, respectively. G tests were positive in all 28 cases of the true positive group, and 4 in 6 non-fungal infection cases. The sensitivity, specificity, positive predictive value and negative predictive value were 100%, 33.3%, 87.5% and 100%, respectively. G test is more sensitive than GM test (P = 0.015), but there was no significant difference in specificity of the two tests (P = 0.242). In 19 of 21 patients with GM test positive, anti-fungal treatment was effective, and GM value gradually decreased to negative, two invalid patients were persistent with GM test positive. After two weeks treatment, the average GM value was significantly lower in the effective group than in the ineffective group (P < 0.05). BG values in the responded patients showed a gradual decline similar to that of GM values, but not to negative. The changes of BG value in ineffective group varied with a trend upward. The changes in BG value had no relation with treatment effectiveness.

CONCLUSIONSSerum GM and BG antigens detection provides strong evidence for early diagnosis of IFI. Combination of GM and G tests can improve the diagnostic specificity and reduce the false positive GM test seems superior to G test for monitoring GM and BG values during treatment.

Adult ; Aged ; Aged, 80 and over ; Antigens, Fungal ; blood ; immunology ; Female ; Hematologic Neoplasms ; immunology ; microbiology ; Humans ; Male ; Mannans ; immunology ; Middle Aged ; Mycoses ; blood ; immunology ; Young Adult ; beta-Glucans ; immunology

Neospora caninum is the etiologic agent of bovine neosporosis, which affects the reproductive performance of cattle worldwide. The transmembrane protein, NcSRS2, and dense-granule protein, NcGRA7, were identified as protective antigens based on their ability to induce significant protective immune responses in murine neosporosis models. In the current study, NcSRS2 and NcGRA7 genes were spliced by overlap-extension PCR in a recombinant adenovirus termed Ad5-NcSRS2-NcGRA 7, expressing the NcSRS2-NcGRA7 gene, and the efficacy was evaluated in mice. The results showed that the titer of the recombinant adenovirus was 10(9)TCID50/ml. Three weeks post-boost immunization (w.p.b.i.), the IgG antibody titer in sera was as high as 1:4,096. IFN-gamma and IL-4 levels were significantly different from the control group (P<0.01). This research established a solid foundation for the development of a recombinant adenovirus vaccine against bovine N. caninum.
Adenoviridae/*genetics ; Animals ; Antibodies, Fungal/blood ; Antigens, Fungal/genetics/*immunology ; *Drug Carriers ; Fungal Proteins/genetics/*immunology ; Fungal Vaccines/administration & dosage/genetics/*immunology ; Immunoglobulin G/blood ; Interferon-gamma/blood ; Interleukin-4/blood ; Mice ; Mice, Inbred BALB C ; Neospora/genetics/*immunology ; Recombinant Fusion Proteins/genetics/immunology ; Vaccines, Synthetic/administration & dosage/genetics/immunology

Oral vaccination may be the most efficient way of inducing an immune response at the remote mucosal site through the common mucosal immune network. Antigenspecific secretory IgA (sIgA) is the major immunoglobulin type generally detected in the secretions of experimental animals following an effective oral immunization. Actinobacillus pleuropneumoniae causing disease in the lung of pig initially interacts, colonizes, and infects the host tissues at the mucosal surface of the respiratory tract. Also, importantly for A. pleuropneumoniae protection, the quantity of sIgA in the lung had merits associated with the mucosal immunity. However, there is no simple method to monitor the level of sIgA as an indicator for the induction of local immune responses by an oral vaccination in the target tissue. Therefore, the relationship between sIgA and IgG was analyzed to evaluate the induction of local immune responses by an oral immunization with Saccharomyces cerevisiae expressing the apxIA and apxIIA genes of A. pleuropneumoniae in this study. The correlation coefficient of determination (r2 x 100) for paired samples in both vaccinated and control groups showed a significant positive-relationship between IgG in sera and sIgA in the lung or intestine. These results indicated that IgG antibody titers in sera could be useful to indirectly predict local immune response, and sIgA, in the lung or intestine to evaluate the efficacy of an oral vaccination.
Actinobacillus pleuropneumoniae ; Administration, Oral ; Animals ; Antigens, Fungal/*immunology ; Bacterial Proteins/genetics/immunology ; Bacterial Vaccines/*immunology ; Disease Models, Animal ; Female ; Hemolysin Proteins ; Immunity, Mucosal/*immunology ; Immunoglobulin A, Secretory/*analysis ; Immunoglobulin G/*blood ; Intestine, Small/immunology ; Lung/immunology ; Mice ; Mice, Inbred BALB C ; Saccharomyces cerevisiae/*immunology

This study was purposed to evaluate the sensitivity and specificity of G/(1, 3-β-D glucan)/GM (galactomannan) combined detection for early diagnosis of invasive fungal disease (IFD), determine the GM positive cut-off value, and investigate the change of diagnostic level before and after G/GM test and the relation of GM value with therapeutic effect. The ELISA with double antibody sandwich was used to detect the serum levels of G and G/M. The results showed that according to determined GM positive cut-off value, the sensitivity, specificity, positive and negative predictive value of G/GM combined detection were 100%, 65.7%, 67.6% and 100%, respectively. The case number of the clinical diagnosis of IFD increased from 1 to 24 cases, the diagnosis of 12 non-infective patients was changed as suspected IFD with the help of G/GM combined detection; the GM cut-off value decreased in patients whose GM cut-off value was higher before therapy and antifungal therapy was effective, while the GM cut-off value increased in patients no-responded to therapy. It is concluded that G/GM combined detection can increased the sensitivity of the diagnosis and reduce false negative rate in the early diagnosis of IFD. The dynamically monitoring G/GM cut-off value can be used as evaluation indicator of therapeutic efficacy.
Adolescent ; Adult ; Aged ; Antigens, Fungal ; analysis ; Child ; Child, Preschool ; Early Diagnosis ; Female ; Hematologic Neoplasms ; complications ; microbiology ; Hematopoietic Stem Cell Transplantation ; Humans ; Male ; Middle Aged ; Mycoses ; diagnosis ; etiology ; immunology ; Sensitivity and Specificity ; Young Adult

OBJECTIVETo explore the relationship between the optical density index of serum aspergillus galactomannan (GM) assay and invasive aspergillosis (IA).

METHODSFrom Jan 2008 to Dec 2011, 825 hematological diseases patients with neutrophil count <0.5×10⁹/L⁹ by continuous blood count tests were admitted into our hospital. The optical density index of GM assay was ≥0.5 at least once. Of 825 patients, 247 cases were manifested as fever during hospitalization. The optical density index of GM antigen was detected by enzyme-linked immunosorbent assay, and the sensitivity and specificity of optical density ranged in 0.5-1.5.

RESULTSIn this study, the sensitivity and specificity of GM assay with continuous twice samples (73% and 93%, respectively) were higher than single sample (66% and 80%, respectively) when optical density index ≥1.0. 69 cases were diagnosed as proven IA with the incidence rate of 8.36%.

CONCLUSIONThe cut-off level for serum GM antigen assay should be decided as optical density index in two continuous samples of ≥1.0.

Adolescent ; Adult ; Aged ; Antigens, Fungal ; blood ; Aspergillosis ; blood ; diagnosis ; etiology ; Enzyme-Linked Immunosorbent Assay ; Female ; Hematologic Diseases ; blood ; microbiology ; Humans ; Male ; Mannans ; blood ; immunology ; Middle Aged ; Sensitivity and Specificity ; Young Adult

INTRODUCTIONInvasive fungal pan-sinusitis can present atypically with severe acute visual loss with minimal anterior orbital inflammation. We describe 2 such cases with a background of uncontrolled diabetes.

CLINICAL PICTURERespective clinical presentations of orbital apex and cavernous sinus syndromes were associated with isolation of Aspergillus galactomannan and Rhizopus.

TREATMENTUrgent extensive surgical debridement and systemic antifungal is necessary.

OUTCOMEClinical improvement of the ocular motor nerves can be expected within 2 months of treatment but visual loss is usually permanent.

CONCLUSIONUnderlying pansinusitis is an important differential for acute visual loss, especially in uncontrolled diabetics. Early treatment determines outcome.

Adult ; Antifungal Agents ; therapeutic use ; Antigens, Fungal ; analysis ; Aspergillosis ; complications ; diagnosis ; therapy ; Aspergillus ; immunology ; isolation & purification ; Debridement ; methods ; Diabetes Complications ; blood ; complications ; therapy ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Mucormycosis ; complications ; diagnosis ; therapy ; Rhizopus ; immunology ; isolation & purification ; Sinusitis ; complications ; diagnosis ; therapy ; Tomography, X-Ray Computed ; Vision, Low ; diagnosis ; etiology ; therapy