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

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

BACKGROUND: Neutrophils fixed on a slide are used in indirect immunofluorescence assay (IIF) for anti-neutrophil cytoplasmic antibody (ANCA). A simple method to prepare ANCA slides is described in this report. METHODS: This method uses an overlay method to separate WBC from EDTA whole blood and Octospottrade mark kits to cytocentrifuge WBC onto 8-well slides. This in-house ANCA IIF was compared with commercial ANCA IIF using 40 sera, and with enzyme linked immunosorbent assay for anti-myeloperoxidase (MPO) antibody (MPO ELISA) using 117 sera. RESULTS: In ANCA IIF parallel tests using 10 positive and 30 negative sera, in-house slides were in good qualitative agreement, 97.5% (39/40) with commercial kits (kappa=0.93). Fluorescence pattern and intensity of positive reaction was also highly concordant between two methods. In MPO ELISA positive cases, the positivity rate of in-house ANCA IIF was 97.8% (45/46), which accounts for its assay sensitivity for the presence of anti-MPO antibody. CONCLUSIONS: In-house ANCA slides was convenient and cost-efficient to be prepared. As these slides were found to have acceptable agreement and sensitivity as compared with commercial kits, in-house ANCA IIF is considered to be useful to screen ANCA in combination with MPO ELISA in routine clinical laboratory practice.
Antibodies, Antineutrophil Cytoplasmic ; Edetic Acid ; Enzyme-Linked Immunosorbent Assay ; Fluorescence ; Fluorescent Antibody Technique, Indirect ; Neutrophils

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

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

The authors investigated the incidence of stratum corneum and upper epidermal cytoplasmic antibodies with 30 untreated and 20 treated psoriasis sera, and normal human sera using normal human skins of 5 different sites and psoriatic lesion by the method of indirect immunofluorescence in order to evaluate immunologic responses i n psoreasis. The results are summarized as follows . I) The positivity of stratum corneum antibodies in untreated psoriasis sera(78.7%) was si, nificantly higher than that in normal sera(64.0%). The incidence of stratum corneum antibodies in untreated psoriasis sera was found to be the highest in the arm, followed by the scalp, leg, abdomen, and face as substrate. 2) The positivity of stratum corneum antibodies in psoriasis and normal human sera was significantly higher when tested with the psoriatic lesion as substrate than norma! skin as substrate, 3) The titer of stratum corneum antibodies in 5 sera using human skin obtained from 5 different sites on the body as substrate are the highest in the arm, and leg, and(in decreasing order of frequency) the scalp, abdomen, and face. 4) The positivity of upper epidermal cytoplhsmic antibodies in normal human sera (40.7%) was significa.ntly higher than that in untreated psoriasis sera(21.3%). 5) Ir.. the majority of cases, upper epidermal cytoplasmic antibodies coexisted with straturn corneum antibodies in the sera of patients with psoriasis and in the sera of normal humans.
Abdomen ; Antibodies* ; Arm ; Cytoplasm* ; Fluorescent Antibody Technique, Indirect ; Humans ; Humans* ; Incidence* ; Leg ; Psoriasis* ; Scalp ; Skin

BACKGROUND: IFN-gamma plays an important role in host response to intracellular organisms such as mycobacterium. Human infection with mycobacterium leads to a wide variety of outcomes, ranging from asymptomatic infection to widespread and rapidly fatal disease. Recent reports suggest that alteration of the function of IFN-gamma caused by a defective IFN-gamma receptor gene can explain different host response to mycobacterium. In this study, we investigated the role of IFN-gamma in the development of chronic refractory tuberculosis. METHODS: The LPS-induced TNF-alpha production with or without IFN-gamma priming was compared by using monocytes taken from recently diagnosed tuberculosis, chronic refractory tuberculosis patients and controls. And the IFN-gamma receptor was measured by indirect fluorescent antibody technique to know whether change in the priming effect of IFN-gamma is related to IFN-gamma receptor deficiency or not. RESULTS: The ratio of TNF-alpha produced in response to stimulation with INF-gamma and LPS to LPS alone was 13.5 +/- 7.6 in controls, 10.8 +/- 6.4 in recently diagnosed tuberculosis patients and 6.7 +/- 3.9 in chronic refractory tuberculosis patients. The priming effect of IFN-gamma significantly decreased in chronic refractory tuberculosis patients compared with that in controls (p=0.002). However, IFN-gamma receptor deficiency was detected in one of chronic refractory tuberculosis patients. CONCLUSION: The decrease of the priming effect of IFN-gamma may play an important role in the development of chronic refractory tuberculosis, and in some patients, this may be related to the IFN-gamma receptor deficiency.
Asymptomatic Infections ; Fluorescent Antibody Technique, Indirect ; Humans ; Monocytes ; Mycobacterium ; Tuberculosis* ; Tumor Necrosis Factor-alpha

In the present study, we have tried to isolate and identify herpes simplex virus type 2(HSV 2) from clinical specirnens, which were inoculated into Vero cell line and grown. Eight strains of viruses were isolated from 20 suspected cases diagnosed from the pr ivate clinics in Seoul. Viruses isolated from 4 rnale and 1 female cases with active lesion were identified to the HSV 2 by indirect immunofluorescence using monoclonal antibody to HSV-2. In addition, morphology of the isolated viruses were observed under electron microscope.
Female ; Fluorescent Antibody Technique, Indirect ; Herpes Simplex* ; Herpesvirus 2, Human* ; Humans ; Seoul ; Simplexvirus* ; Vero Cells