Indirect fluorescent antibody tests were performed with sera of paragonimiasis patients and skin test positive sera against Paragonimus antigen. Paragonimus antigen was prepared from lyophilized adult worms of P. westermani by defatting with ethyl-ether before extracting with barbital buffered saline. Preparation of Paragonimus antigen for the indirect fluorescent antibody test was based upon Sato's method used for sero-diagnosis of anisakiasis, with Sephadex G-25 instead of Sepharose 4B. The results were as follows: The indirect fluorescent antibody titers of paragonimiasis patient's sera ranged from 1:64 to 1:512, whereas the control sera showed titers of less than 1:16. As controls, Clonorchis patient's sera and parasite-free healthy human sera were used. In indirect fluorescent antibody tests, the skin test positive human sera against Paragonimus antigen showed a positive rate of 41.5 per cent in the case of titers more than 1:40. On the other hand, complement fixation tests on the same sera showed a positive rate of 32.5 per cent in the case of titers more than 1:20.
parasitology-helminth-trematoda ; paragonimiasis ; Paragonimus westermani ; diagnosis ; indirect fluorescent antibody tests ; serum ; ethyl-ether

Autoimmune neutropenia of infancy (AIN) is caused by increased peripheral destruction of neutrophils as a result of antibodies in patients' blood that are directed against their own neutrophils. Due to non-specific symptoms, benign clinical courses, and cumbersome diagnostic tests, AIN are commonly undetected. Antineutrophil antibody test for diagnosis of AIN has recently become available. Compared to its relatively lower absolute neutrophil count (ANC), the clinical course of AIN is mostly benign. Therefore, although treatment is not usually necessary for AIN, it is applicable in order to rule out other significant diseases, such as severe congenital neutropenia (SCN), which can be transformed to myelodysplastic syndrome or acute myelocytic leukemia. For this reason, several treatments can be used for neutropenia: granulocyte-colony stimulating factor (G-CSF) for SCN, trimethoprim and sulfamethoxazole (TMP-SMX) for prophylaxis. Here we report on two cases of AIN confirmed by indirect immunofluorescence test using flow cytometry.
Antibodies ; Diagnostic Tests, Routine ; Flow Cytometry ; Fluorescent Antibody Technique, Indirect ; Leukemia, Myeloid, Acute ; Myelodysplastic Syndromes ; Neutropenia ; Neutrophils ; Sulfamethoxazole ; Trimethoprim

BACKGROUND: Anti-mitochondrial antibodies (AMA) are a hallmark of primary biliary cirrhosis (PBC); however, low titers of AMA are also detected in some patients without PBC. We evaluated the clinical value of commercial rat kidney/stomach sections and an additional biochip coated with mitochondrial antigen M2 (pyruvate dehydrogenase complex). METHODS: A total of 124 patients who had been tested for AMA were evaluated. Results of AMA, antibodies to M2, and antinuclear antibody were reviewed retrospectively and searched for clinical and laboratory data to diagnose PBC. AMA and M2 antibody were assayed by an indirect immunofluorescence assay using EUROPLUS kit (Euroimmun, Luebeck, Germany). RESULTS: In 10 of the 124 patients, a diagnosis of PBC was established by AMA, liver function test or liver biopsy. The sensitivity and specificity of rat kidney/stomach section, M2 antibody, and coarse cytoplasmic fluorescent pattern of HEp-2 cell were 80.0, 75.0, 88.9% and 97.4, 98.2, 97.3%, respectively; however, these differences were not statistically significant. Six patients with coarse cytoplasmic pattern of HEp-2 cell at 1: 320 dilution were positive for both rat kidney/stomach sec-tion and M2 antibody. Two of five patients with coarse cytoplasmic pattern at below 1: 80 dilution were diagnosed as PBC, yet all of them were negative for M2 antibody. CONCLUSIONS: M2 biochip test would be convenient to test simultaneously with rat kidney/stomach section and it provided results similar to those of the preexisting serological tests for PBC.
Animals ; Antibodies ; Antibodies, Antinuclear ; Biopsy ; Cytoplasm ; Diagnosis ; Fluorescent Antibody Technique, Indirect ; Humans ; Liver ; Liver Cirrhosis, Biliary ; Liver Function Tests ; Oxidoreductases ; Rats ; Retrospective Studies ; Sensitivity and Specificity ; Serologic Tests

OBJECTIVES: Q fever is a zoonotic disease that occurs worldwide; however, little is known about its prevalence in South Korea. We attempted to determine the prevalence of Q fever seroreactivity among Korean slaughterhouse workers and the risk factors for seroreactivity according to the type of work. METHODS: The study was conducted among 1503 workers at a total of 73 slaughterhouses and 62 residual-product disposal plants. During the study period, sites were visited and surveys were administered to employees involved in slaughterhouse work, and serological tests were performed on blood samples by indirect immunofluorescence assays. Serological samples were grouped by job classification into those of slaughter workers, residual-product handlers, inspectors and inspection assistants, and grading testers and testing assistants. Employee risk factors were analyzed according to the type of work. RESULTS: Out of 1481 study subjects who provided a blood sample, 151 (10.2%) showed reactive antibodies. When these results were analyzed in accordance with the type of work, the result of slaughter workers (11.3%) was similar to the result of residual-product handlers (11.4%), and the result of inspectors and assistants (5.3%) was similar to the result of grading testers and assistants (5.4%). Among those who answered in the affirmative to the survey question, “Has there been frequent contact between cattle blood and your mouth while working?” the proportions were 13.4 and 4.6%, respectively, and this was identified as a risk factor that significantly varied between job categories among slaughterhouse workers. CONCLUSIONS: This study found a Q fever seroreactivity rate of 10.2% for slaughterhouse workers, who are known to be a high-risk population. Contact with cattle blood around the mouth while working was the differential risk factor between job categories among slaughterhouse workers.
Abattoirs* ; Animals ; Antibodies ; Cattle ; Classification ; Coxiella burnetii ; Fluorescent Antibody Technique, Indirect ; Korea* ; Mouth ; Prevalence ; Q Fever* ; Risk Factors ; Serologic Tests ; Zoonoses

BACKGROUND: Despite rigorous investigations, the etiology of community-acquired pneumonia remains unknown in about 50% of hospitalized patients. The diagnosis of the etiological agent is becoming more challenging and more critical as number of newer pathogens have been recognized in recent years. In the 3-year period prospective study we investigated adult patients with community-acquired pneumonia for Legionella, Leptospira, Hantaan virus and Orientia tsutsugamushi as potential etiologic agents. METHODS: A prospective multicenter study was performed from May 1997 to April 2000. A total of 431 patients with community-acquired pneumonia under the inclusion criteria were examined for specific microbial diagnosis; sputum culture and PCR, and serologic teats including indirect immunofluorescence antibody (IFA) test for Legionella, and hemagglutination tests for Leptospira, Hantaan virus and O. tsutsugamushi. Etiologic diagnosis was determined on the basis of the review of case record forms and specific laboratory diagnostic criteria. RESULT: During the study period a total of 385 sputum and 283 serum samples were examined. Legionella pneumonia was diagnosed in 2.3% (10/431) of the cases examined: 1.4% cases with PCR-positive (5/ 367) and 2.1% with positive IFA test (6/283). Leptospirosis and scrub typhus were diagnosed in 0.4% (1/ 252) and 2.0% (5/252), respectively. All 5 cases diagnosed as scrub typhus occurred in late fall, and rash or eschar was not found. None of cases was Hantaan virus infection. CONCLUSION: The results suggest that Legionella, Leptospira, and O. tsutsugamushi should be considered in the etiologic diagnosis and in empirical antibiotic therapy of community-acquired pneumonia.
Adult* ; Diagnosis ; Exanthema ; Fluorescent Antibody Technique, Indirect ; Hantaan virus* ; Hemagglutination Tests ; Humans ; Legionella* ; Leptospira* ; Leptospirosis ; Orientia tsutsugamushi* ; Pneumonia* ; Polymerase Chain Reaction ; Prospective Studies* ; Scrub Typhus ; Sputum

BACKGROUND: Serologic test is important in the evaluation of patients with connective tissue diseases(CTD). It does help to confirm a clinical diagnosis, classify subsets of CTD and predict prognosis. In Korean patients with various CTD, the positive rate of each autoantibody has not been reported in dermatologic literatures. OBJECTIVES: The purpose of this study was aimed at investigating the positive rate of autoantibodies in Korean patients with CTD. PATIENTS AND METHODS: The 173 patients who visited the dermatologic clinic of Asan medical center from 1995 to 2000 were diagnosed as having systemic lupus erythematosus(SLE), discoid lupus erythematosus(DLE), dermatomyositis or scleroderma. Fifty eight patients of SLE, 35 patients of DLE, 34 patients of dermatomyositis and 46 patients of scleroderma were enrolled. Using the sera from the total 173 patients, the presence of antinuclear antibody(ANA) was detected by an indirect immunofluorescence test using cultured human cell substrates(HEp-2 cells). The presence of extractable nuclear antigen(RNP, Sm, Ro(SS-A), La(SS-B), Scl-70, Jo-1) was detected by gel double immunodiffusion. Anti-double-stranded DNA(dsDNA) was detected by radioimmunoassay. RESULTS: 1. Of the 58 patients with SLE, the sera of 56 patients(96%) was positive to ANA. Thirty four patients(58%) had anti-dsDNA antibodies. Thirteen patients(22%) had anti-RNP antibodies. Twenty nine patients(50%) had anti-Ro(SS-A) antibodies while 5 patients(8%) had anti-La(SS-B) antibodies. Only 5 patients(8%) had anti-Sm antibodies. 2. Of the 35 patients with DLE, the sera of 17 patients(48%) was positive to ANA. Among 10 patients tested, only one patient(10%) had anti-RNP antibodies. Two patients(20%) had both anti-Ro(SS-A) and anti-La(SS-B) antibodies. 3. Of the 34 patients with Dermatomyositis, the sera of 16 patients(47%) was positive to ANA. Among 13 patients tested, one patient(7%) had anti-dsDNA antibody. Among 20 patients tested, two patients(10%) had anti-Ro(SS-A) antibodies. Two patients(10%) had anti-Jo-1 antibodies. 4. Of the 46 patients with scleroderma, the sera of 24 patients(52%) was positive to ANA. Among 12 patients tested, two patients(16%) had anti-dsDNA antibodies. Among 30 patients tested, nine patients(30%) had anti-Scl-70 antibodies. CONCLUSIONS: 1. In Korean patients with CTD, the positive rate of various autoantibodies were similar to the previous western reports except some minor differences; (1) lower positive rate of anti-Sm and higher positive rate of anti-Ro(SS-A) in SLE. (2) lower positive rate of anti-Jo-1 in dermatomyositis. (3) higher positive rate of anti-Scl-70 in scleroderma. 2. SLE patients have significantly higher percentage of ANA, anti-dsDNA, anti-RNP, anti-Sm, anti-Ro(SS-A) antibodies in compared to DLE, dermatomyositis, and scleroderma patients.
Antibodies ; Autoantibodies* ; Chungcheongnam-do ; Connective Tissue Diseases* ; Connective Tissue* ; Dermatomyositis ; Diagnosis ; Fluorescent Antibody Technique, Indirect ; Humans ; Immunodiffusion ; Prognosis ; Radioimmunoassay ; Serologic Tests

OBJECTIVETo compare the three Anti-dsDNA antibody detecting test (IIF, ELISA, Farr) with 200 serum samples to evaluate which one has higher sensitivity and specificity.

METHODS200 serum samples including 120 serum samples of SLE, 20 serum samples of rheumatoid arthritis, 20 serum samples of MCTD, 20 serum samples of SS, 20 serum samples of PSS and 50 serum samples of healthy measured by IIF, Farr and ELISA.

RESULTSDetection the Anti-dsDNA antibody of the serum sample with the methods of IIF, ELISA and Farr. The positive percentage of Anti-dsDNA in SLE is 25%, 32% and 32%, while in RA is 0, 0 and 0; in PSS is 0, 0 and 5%; in SS is 0, 0 and 0; in healthy is 0, 0 and 0.

CONCLUSIONDetection the Anti-dsDNA antibody with two method in the same time, especially with IIF and ELISA, will heighten the positive rate than with single method and will be helpful for the diagnosis of SLE.

Antibodies, Antinuclear ; analysis ; immunology ; Case-Control Studies ; DNA ; immunology ; Enzyme-Linked Immunosorbent Assay ; Fluorescent Antibody Technique, Indirect ; Humans ; Immunologic Tests ; methods ; Lupus Erythematosus, Systemic ; diagnosis ; immunology ; Radioimmunoassay

OBJECTIVETo establish a method of indirect immunofluorescence (IIF) to measure DNA (mDNA)-associated autoantibodies to cell membrane, and to evaluate diagnostic value of the anti-mDNA antibodies in patients with juvenile systemic lupus erythematosus (SLE) in comparison with anti-dsDNA antibody.

METHODSForty-four children with SLE were enrolled in this study. As a control group, 30 children with other rheumatic diseases were also enrolled. Anti-mDNA and anti-dsDNA antibodies were measured by IIF. Anti-smooth muscle (Sm) antibodies were measured by immuno-double diffusion (ID) and IIF.

RESULTSOut of 44 juvenile SLE patients, 34 (77.27%) were seropositive for anti-mDNA, which was significantly higher than that of patients with other rheumatic diseases (20.00%, P<0.05). The sensitivity and specificity of anti-mDNA for juvenile SLE diagnosis were 77.27% and 80.00%, respectively. The positive predictive value and negative predictive value were 85.00% and 70.59%, respectively. The positive rate of anti-mDNA in SLE lacking of anti-dsDNA and anti-Sm antibodies were 68.00% (17/25) and 79.49% (31/39), respectively.

CONCLUSIONThe detection of anti-mDNA antibodies is useful for diagnosis of juvenile SLE, especially in patients who are negative for anti-dsDNA antibodies and anti-Sm antibodies.

Adolescent ; Antibodies, Antinuclear ; analysis ; Case-Control Studies ; Cell Membrane ; immunology ; Child ; Female ; Fluorescent Antibody Technique, Indirect ; methods ; Humans ; Lupus Erythematosus, Systemic ; immunology ; Male ; Predictive Value of Tests ; Sensitivity and Specificity

Severe fever with thrombocytopenia syndrome (SFTS) is caused by the SFTS virus (SFTSV). The SFTSV appears to have a wide host range, as SFTSV-positive ticks have been isolated from both farm animals and wild rodents. Therefore, it is important to monitor SFTSV-positive animals to prevent the transmission of SFTSV from animals to humans. Previously, we developed a competitive enzyme-linked immunosorbent assay (cELISA) to detect SFTSV-specific antibodies from field animals and compared the cELISA results to those from an indirect immunofluorescence assay (IFA). In this study, cELISA results were compared to and evaluated against the results from both an IFA and a virus neutralization (VN) test of 193 bovine serum samples (including two bovine positive control sera) and 70 horse serum samples. The consistency (98.9%) between cELISA and VN results was higher than that (97.4%) between cELISA and IFA for the bovine serum samples. Similarly, for the horse serum samples, the consistency (88.6%) between cELISA and VN results was higher than that (84.3%) between the cELISA and IFA. These findings indicate that our newly developed cELISA can be used for surveillance or epidemiological studies of SFTSV in animals.
Animals ; Animals, Domestic ; Antibodies ; Enzyme-Linked Immunosorbent Assay* ; Epidemiologic Studies ; Fever* ; Fluorescent Antibody Technique, Indirect ; Horses ; Host Specificity ; Humans ; Neutralization Tests* ; Rodentia ; Thrombocytopenia* ; Ticks

We report a rare case of bullous scabies with bullous pemphigoid in a 59-year-old male patient. He presented with a 9-month history of exhibiting multiple, variable-sized, red-to-brown, pruritic cutaneous patches, and papules with tense bullae on his whole body. A direct smear of the bullous lesions was performed and was negative for scabies mites. Histopathologic findings (hematoxylin and eosin staining) revealed Sarcoptes scabiei in the stratum corneum. There were sub-epidermal blisters with massive eosinophil and lymphocyte infiltration in the epidermis and upper dermis. Direct immunofluorescence microscopy showed linear deposition of IgG at the dermo-epidermal junction. Indirect immunofluorescence microscopy of samples acquired for the salt-split skin test showed linear IgG deposition in the epidermis. Skin lesions improved after the patient was treated with an anti-scabietic and steroids.
Blister ; Dermis ; Eosine Yellowish-(YS) ; Eosinophils ; Epidermis ; Fluorescent Antibody Technique, Direct ; Fluorescent Antibody Technique, Indirect ; Humans ; Immunoglobulin G ; Lymphocytes ; Male ; Microscopy ; Middle Aged ; Mites ; Pemphigoid, Bullous* ; Sarcoptes scabiei ; Scabies* ; Skin ; Skin Tests ; Steroids ; Transcutaneous Electric Nerve Stimulation