No abstract available.
Chlamydia* ; Fluorescent Antibody Technique, Indirect*

No abstract available.
Diagnosis* ; Fluorescent Antibody Technique, Indirect* ; Herpes Simplex* ; Simplexvirus*

The pathophysiology of glandular dysfunction in Sjögren's syndrome (SS) has not been fully elucidated. Previously, we reported the presence of autoantibodies to AQP-5 in patients with SS, which was associated with a low resting salivary flow. The purpose of this study was to investigate the presence of anti-AQP1 autoantibodies. To detect anti-AQP1 autoantibodies, cell-based indirect immunofluorescence assay was developed using MDCK cells that overexpressed human AQP1. By screening 112 SS and 52 control sera, anti-AQP1 autoantibodies were detected in 27.7% of the SS but in none of the control sera. Interestingly, the sera that were positive for anti-AQP1 autoantibodies also contained anti-AQP5 autoantibodies in the previous study. Different from anti-AQP5 autoantibodies, the presence of anti-AQP1 autoantibodies was not associated with the salivary flow rate. Although anti-AQP1 autoantibodies are not useful as a diagnostic marker, the presence of autoantibodies to AQP1 may be an obstacle to AQP1 gene therapy for SS.
Aquaporin 1 ; Autoantibodies* ; Fluorescent Antibody Technique ; Fluorescent Antibody Technique, Indirect ; Genetic Therapy ; Humans ; Madin Darby Canine Kidney Cells ; Mass Screening

The distribution of pemphigus subclass autoantibodies in a patient with pemphigus vulgaris (PV) has been investigated by semiquantitative indirect immunofluorescence (IIF), using the HP series monoclonal antibodies specific for four human IgG subclasses on human foreskins. IgG1 and IgG4 intercellular substance-specific autoantibodies were detected in the serum of the patient, whereas IgG2 and IgG3 autoantibodies were absent. In addition to foreskins, human tonsillar epithelia were used as substrates of IIF for detecting the PV autoantibodies and it was one of satisfactory substitutes for monkey esophagus.
Antibodies, Monoclonal ; Autoantibodies* ; Esophagus ; Fluorescent Antibody Technique, Indirect ; Foreskin ; Haplorhini ; Humans ; Immunoglobulin G* ; Pemphigus*

OBJECTIVE: To evaluate the associations between clinical and laboratory parameters and antinuclear antibodies (ANA)in rheumatoid arthritis (RA). METHODS: We determined functional status,disease duration,hemoglobin,platelet count,erythrocyte sedimentation rate,C-reactive protein,rheumatoid factor (RF),ANA,and antiperinuclear factor in 89 RA patients.ANAs were studied by indirect immunofluorescence using Hep-2 cells.The medical records of the patients were reviewed. RESULTS: ANAs were detected in 30.3%of RA patients and 66.6%of ANAs were below the titer of 1:80.Nine patients had ANAs above the titer of 1:160, five of them had additional diseases besides RA.The ANA positivity was correlated only with the presence of RF.Two of the 5 ANA-positive patients without RF had thyroid disease. CONCLUSIONS: Except for RF,no significant correlation was observed between clinical and laboratory parameters and ANA positivity,but ANA positivity appears to be associated with the general autoimmunity.It is needed to search other associated pathologic conditions for ANA positive RA patients without RF.
Antibodies, Antinuclear* ; Arthritis, Rheumatoid* ; Fluorescent Antibody Technique, Indirect ; Humans ; Medical Records ; Rheumatoid Factor ; Thyroid Diseases

BACKGROUND: The gold standard for antinuclear antibody (ANA) screening is the indirect immunofluorescence (IIF) assay with human epithelial cells (HEp-2). However, a number of substantial disadvantages of manual IIF assays have highlighted the need for the automation and standardization of fluorescent ANA (FANA) testing. We evaluated the performance of EUROPattern Suite (Euroimmun AG, Germany), an automated FANA image analyzer, with regard to ANA detection and pattern recognition compared with conventional manual interpretation using the fluorescence microscopic IIF assay. METHODS: A total of 104 samples including 70 ANA-positive sera and 34 ANA-negative sera collected from September to October 2015 were included. The sensitivity, specificity, and pattern recognition function were evaluated to determine the performance of EUROPattern Suite compared with the manual IIF assay results. RESULTS: The sensitivity and specificity of EUROPattern Suite for ANA detection were 94.3% and 94.1%, respectively. The concordance rate between the two methods was 94.2%. For pattern recognition, 45.7% of the samples were assigned identical ANA patterns including simple and mixed. When major pattern matching was considered, 83.7% (41/49) and 95.2% (20/21) of the samples with simple and mixed patterns, respectively, showed concordant results between the two methods. CONCLUSIONS: EUROPattern Suite, an automated FANA image analyzer, provides a viable option for distinguishing between positive and negative results, although the ability to assign specific patterns is insufficient to replace manual microscopic interpretation. This automated system may increase efficiency in laboratories, in which a large number of samples need to be processed.
Antibodies, Antinuclear* ; Automation ; Epithelial Cells ; Fluorescence* ; Fluorescent Antibody Technique, Indirect ; Humans ; Mass Screening ; Sensitivity and Specificity

No abstract available.
Adult ; Cornea/*metabolism ; Fluorescent Antibody Technique, Indirect ; Humans ; Microscopy, Fluorescence ; Middle Aged ; alpha-Synuclein/*metabolism

Group A rotaviruses are the most common causes of gastroenteritis among infants and young children. The outer capsid layer of the virus is composed of two structural proteins, VP4 and VP7, and they play important roles in protection by eliciting neutralization antibodies. Group A rotaviruses are subdivided into distinct G and P serotypes according to the antigenic differences of the VP7 and VP4, respectively. Rotavirus G9 serotype was thought to be the fifth most common serotype circulating among the population worldwide. In this study, G9 human rotaviruses (HRV) were isolated from fecal samples using MA104 cells and characterized. Characteristic cytopathic effects of rotavirus were observed and rotaviral antigens were confirmed by indirect immunofluorescence antibody test in MA104 cells inoculated with isolated HRV strains. The nucleotide sequences of the VP7 gene of Korean G9 HRV isolated in this study were determined and compared with those of other recent and prototype G9 rotavirus strains from other parts of the world. Also, the nucleotide sequences of VP4 and NSP4 gene of Korean G9 HRV were determined and compared with those of other rotavirus strains from other countries. The results showed that the Korean HRV isolates belong to a G9, P[8] and NSP4 B genotype. The Korean G9 HRV isolates and their nucleotide sequence data would be usefully applied for the vaccine development of HRV in the near future.
Antibodies ; Base Sequence ; Capsid ; Child ; Fluorescent Antibody Technique, Indirect ; Gastroenteritis ; Genotype ; Humans* ; Infant ; Rotavirus*

BACKGROUND: Immunolcgical assays are required for the accurate diagnosis of autoimmune bullous dermatoses including pemphigus vulgaris(PV) and pemphigus foliaceus(PF). In the detection of circulating autoantibodies to pemphigus antigens(desmosomal components), the priority remains controversial between indirect immunofluorescence(IF) and immunoblot(IB) assay. OBJECTIVE: In the present study we compared the sensitivity of indirect IF and that of IB using amplified alkaline phosphatase staining system in the detection of pemphigus autoantibodies. PATIENTS: We selected eight patients with serum endpoint titer of 1:80 in preliminary study. Among these patients three were PV and five were PF. METHODS/RESULTS: The titers of IgG autoantibodies found on indirect IF were confirmed as 1: 80 in all patients, whereas the titers examined by IB assay were much higher, 1: 640 to 1: 2560. In the 3 sera of PV patients, the titers of two cases were 1: 1280 and the third case was 1: 2560. In 5 cases of PF, one was 1:640, two were 1: 1280, and two were 1:2560. CONCLUSION: This result suggests that the immunoblot examination using amplified alkaline phosphatase staining system demonstrates higher sensitivity compared with indirect IF(p=0.0003 by Mann-Whitney U test) in the detection of pemphigus autoantibodies.
Alkaline Phosphatase ; Autoantibodies* ; Diagnosis ; Fluorescent Antibody Technique, Indirect* ; Humans ; Immunoglobulin G ; Pemphigus* ; Skin Diseases, Vesiculobullous

BACKGROUND: Immunolcgical assays are required for the accurate diagnosis of autoimmune bullous dermatoses including pemphigus vulgaris(PV) and pemphigus foliaceus(PF). In the detection of circulating autoantibodies to pemphigus antigens(desmosomal components), the priority remains controversial between indirect immunofluorescence(IF) and immunoblot(IB) assay. OBJECTIVE: In the present study we compared the sensitivity of indirect IF and that of IB using amplified alkaline phosphatase staining system in the detection of pemphigus autoantibodies. PATIENTS: We selected eight patients with serum endpoint titer of 1:80 in preliminary study. Among these patients three were PV and five were PF. METHODS/RESULTS: The titers of IgG autoantibodies found on indirect IF were confirmed as 1: 80 in all patients, whereas the titers examined by IB assay were much higher, 1: 640 to 1: 2560. In the 3 sera of PV patients, the titers of two cases were 1: 1280 and the third case was 1: 2560. In 5 cases of PF, one was 1:640, two were 1: 1280, and two were 1:2560. CONCLUSION: This result suggests that the immunoblot examination using amplified alkaline phosphatase staining system demonstrates higher sensitivity compared with indirect IF(p=0.0003 by Mann-Whitney U test) in the detection of pemphigus autoantibodies.
Alkaline Phosphatase ; Autoantibodies* ; Diagnosis ; Fluorescent Antibody Technique, Indirect* ; Humans ; Immunoglobulin G ; Pemphigus* ; Skin Diseases, Vesiculobullous