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

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

OBJECTIVES: The purpose of this study is to compare newly developed assay for identification of ENA antibody, Phadia EliA ENA with Euroimmun line immunoassay by analyzing the degree of agreement and the individual antibodies between two methods. METHODS: A total of 82 patient samples were used. Indirect immunofluorescence assay using Hep-2 cell was performed to screen the antinuclear antibody (ANA). Euroimmun line immunoassay (LIA) and Phadia EliA ENA assay were tested to identify the antibodies against extractable nuclear antigens (ENAs). Kappa statistics was used to evaluate the degree of agreement. RESULTS: Mean age of patients was 41.0 (8-79), and the M:F ratio was 21:61. ANA was positive in 74 samples, and negative were 8 samples. Kappa analysis of the 82 tested samples showed a moderate strength of agreement (kappa = 0.521, P = 0.000). There were differences in the order of identified individual antibodies between two methods (Ro > La = RNP > Centromere > Sm > Scl-70 in Phadia EliA ENA, Ro > RNP > Sm>La > Scl-70 > Centromere=Jo-1 in Euroimmun LIA). Ro antibody was most frequently identified in Phadia EliA ENA negative-Euroimmun LIA positive specimens (Ro > RNP = Jo-1 > La = Sm = Centromere > Scl-70). CONCLUSIONS: A moderate strength of agreement was observed between the Phadia EliA ENA and the Euroimmun LIA. There seemed to be a significant difference in the ratio of individual antibodies, especially in the anti-Ro and Sm antibodies.
Antibodies ; Antibodies, Antinuclear ; Antigens, Nuclear ; Centromere ; Fluorescent Antibody Technique, Indirect ; Humans ; Immunoassay

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: 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

Type VII collagen is one of the major structural components of the corneal epithelial adhesion complex. Using the immunogold technique combined with indirect immunofluorescence analysis, the fine structural distribution of type VII collagen was studied in the corneas obtained from 5 enucleated hyman eyes (age range, 1-77 years) including one pathologic cornea from graft rejection. The findings on normal cornea corroborated the results from previous studies. In pathologic cornea from graft rejection, type VII collagen antibodies generated linear and irregular patchy fluorescence staining along the epithelial-stromal interface and immunogold binding to type VII collagen mainly occurred within the undulating lamina densa, more densealy distributed anchoring plaques and anchoring fibrils. The distribution of type VII collagen in pathologic human cornea from graft rejection is similar to normal human cornea. But, in pathologic cornea, type VII collagen is more densely distributed in superficial stroma and forms more extended anchoring network, which may be derived from the increased secretion of the type VII collagen due to the activated basal epithelial cell during healing process.
Antibodies ; Collagen Type VII* ; Cornea* ; Epithelial Cells ; Fluorescence ; Fluorescent Antibody Technique, Indirect ; Graft Rejection ; Humans ; Immunohistochemistry

OBJECTIVETo compare indirect immunofluorescence assay (IIFA) and enzyme-linked immunosorbent assay (ELISA) for detecting antinuclear antibodies (ANA) and anti-double-stranded DNA antibodies (anti-dsDNA).

METHODSA total of 125 serum samples were obtained from patients with established or suspected autoimmune disease, and 82 samples were used for ANA detection and 57 for anti-dsDNA detection using both IIFA and ELISA. Fourteen samples were examined for both ANA and anti-dsDNA. In cases where discrepancy occurred in the results by the two methods, extractable nuclear antigens were detected using immunoblotting.

RESULTSThe positivity rate of ANA detected by IIFA and ELISA was significantly different (87.8% and 73.17%, respectively, P<0.01), but the positivity rate of anti-dsDNA was similar between IIFA and ELISA (77.19% and 71.93%, respectively, P>0.05). The percent agreement between the two testing methods with different cutoff values of ANA and anti-dsDNA showed significant differences (P<0.01), and for some uncommon patterns, the percent agreement of the two methods was lowered in ANA detection but remained unchanged for anti-dsDNA with different ANA patterns. High percent agreements of the two methods were obtained with the cutoff ANA titer of 1:100 and the cutoff anti-dsDNA value of weak positivity, but they demonstrated a significant difference in testing low-titer ANA and anti-dsDNA.

CONCLUSIONIIFA is more sensitive than ELISA in detecting the total ANA and anti-dsDNA. ELISA prescreening combined with IIFA can obtain the information of the nuclear pattern and allow the observation of the titer alterations. The combination of two or more testing methods can greatly enhance the accuracy of the results.