No abstract available.
Immunoenzyme Techniques*

No abstract available.
Fluoroimmunoassay* ; Immunoenzyme Techniques*

338 subjects at the age of 17-49 were undergone EIA test. Results showed an increasing trend in Chlamydia trachomatis infection. Clinical manifestations were not detected. The incidence is highest at the age of 17-26 (14.68%), 17-36 (10.67%). In the groups of genital tract inflamation, of uterine neck, of vaginitis the incidence reaches 2.7%, 12.71% and 12.50% respectively
Chlamydia trachomatis ; Immunoenzyme Techniques ; Women

No abstract available.
Immunoenzyme Techniques* ; Mycobacterium tuberculosis* ; Mycobacterium*

PURPOSE: Enzyme immunoassay (EIA) is now a widely used technique. We have described the application of enzyme amplification (sensitive ELISA) in the field of immunoassays of pediatric population. There are two issues with the sensitive ELISA. First is that one can minimize the serum volume, an important concern for pediatricians. The second is the problem of background signal. We demonstrate that it is possible to develop EIAs of high sensitivity and detectability with using very small volume of infant's sera immunized with Hib-PRP vaccine. METHODS: Monoclonal Abs HG11, HP6016, HK2, and KL1 specific for human IgG1, IgG2, C , and C were used. The mAb OAK-1 specific for a subfamily of V I L chains (V Ia), the mAbs KB13 and B12 specific for human V II and V III L chains were also used respectively. Adults were immunized with Hib-CRM vaccine. Immune serum was obtained 4 to 8 wk after immunization. Twenty infants received Hib-CRM vaccine at 2, 4, and 6 month of age and blood samples were obtained at 7 month old. The amount of anti-Hib-PS Ab expressing a V subgroup or V was determined by sandwich type immunoassays using conventional substrate. The amount of the enzyme immobilized to the well was determined with para-nitrophenyl phosphate substrate. A standard ELISA was performed but different substrate (lyophilized NADPH) and amplifier (alcohol dehydrogenase and diaphorase) were used to develop color in final step for enzyme amplification method. RESULTS: We get the dose-response curves obtained using the conventional and amplified detection methods in the anti-PRP Ab assay. The sensitivities of the two assay methods were compared. We can increase the sensitivities four to sixteen folds and minimize the infant's sera volume to perform varing anti-PRP antibody assays. To obtain the advantages of increased sensitivity, any background is minimized by using noncontaminated reagents. CONCLUSIONS: It is possible to develop EIAs of high sensitivity and detectability with using very small volume of infant's sera with using enzyme amplification system (sensitive ELISA).
Adult ; Antibodies* ; Child* ; Enzyme-Linked Immunosorbent Assay* ; Humans ; Immunization ; Immunoassay ; Immunoenzyme Techniques ; Immunoglobulin G ; Indicators and Reagents ; Infant ; Oxidoreductases

BACKGROUND: Hepatitis C virus (HCV) is a major agent of transfusion-associated non-A, non-B hepatitis. The best method for prevention of HCV infection through transfusion is blood donor screening using reliable diagnostic tools. Enzyme immunoassays (EIA) for detection of HCV antibody were developed, but it required equipment and time consuming. In emergency situation such as massive bleeding, HCV screening of blood donor needs more simple, rapid and reliable method. Recently, for rapid detection of HCV antibody, Genedia HCV Rapid assay was developed. To evaluate the usefulness of this assay, comparative studies with third generation anti- HCV EIA and two HCV confirmatory tests (Genedia HCV Confirm 4.0 and Chiron RIBA HCV 3.0 SIA) were performed. METHODS: A total of 156 sera (106 positive and 50 negative), screened by second generation IMx anti-HCV assay (Abbott 2.0; Abbott Laboratories, U.S.A.), were examined with Genedia HCV Rapid (Green Cross, Korea) and Genedia HCV ELISA 3.0 (Green Cross, Korea). The discrepant sera between Genedia HCV Rapid and Genedia HCV ELISA 3.0 were confirmed by both Genedia HCV Confirm 4.0 and Chiron RIBA HCV 3.0 SIA. RESULTS: The concordance rates of Abbott 2.0 vs Genedia HCV Rapid, Abbott 2.0 vs Genedia HCV ELISA 3.0, and Genedia HCV Rapid vs Genedia HCV ELISA 3.0 were 88.4%, 89.7%, and 96.1%, respectively. The sensitivity, specificity, positive predictive value and negative predictive value of Genedia HCV Rapid for Genedia HCV Confirm 4.0 were 97.7%, 100%, 98.9%, and 97.0%, respectively, and for Chiron RIBA HCV 3.0 SIA 97.8%, 98.5%, 98.9%, and 97.0%, respectively. Of discrepant 6 sera between Genedia HCV Rapid and Genedia HCV ELISA 3.0, 2 were positive by Genedia HCV Confirm 4.0, and 3 positive by Chiron RIBA HCV 3.0 SIA. However, 14 negative sera by both Genedia HCV rapid and Genedia HCV ELISA 3.0, which were all positive by Abbott 2.0, were all negative by two confirmatory tests. CONCLUSIONS: These data show that Genedia HCV Rapid could be used in emergency blood donor screening for HCV antibody detection.
Blood Donors ; Emergencies ; Enzyme-Linked Immunosorbent Assay ; Hemorrhage ; Hepacivirus* ; Hepatitis C* ; Hepatitis* ; Humans ; Immunoenzyme Techniques ; Mass Screening ; Sensitivity and Specificity

BACKGROUND: Rheumatoid arthritis (RA) is the most common autoimmune rheumatic disease, but sensitive and specific test for its diagnosis is lack. This study evaluated the analytical performance and diagnostic role of a new automated ELISA anti-cyclic citrullinated peptide (anti-CCP) antibody test. METHODS: Anti-CCP antibody test was done with the enzyme-linked immunosorbent assay (ELISA) in serum samples from 49 RA patients and 104 non-RA patients, and 51 healthy subjects. Serum pools were used to determine its precision and linearity. The optimal cut-off values were determined by the receiver-operator characteristics (ROC) curve. The rheumatoid factor (RF) by turbidimetry was also assayed in every samle and the results were compared to anti-CCP for sensitivity and specificity. RESULTS: The total imprecision (CV%) was 4.8%, 7.6% for serum pools with low (mean concentration: 2.7 U/mL) and high (mean concentration :82.2 U/mL) concentration, respectively. Linearity data were acceptable (R2=0.9907). At each optimal cut-off value, the sensitivity of anti-CCP was higher than that of RF (81.6 % vs 69.4%), but statistical significance was not defined. Specificity of anti-CCP was higher than that of RF (95.5% vs 75.5%, p<0.001). A combination of anti-CCP and RF increased sensitivity and specificity to 87.7%, 98.0%, respectively. Nine of 15 (60.0%) sera from RF negative RA patients were positive for anti-CCP. CONCLUSIONS: Anti-CCP ELISA antibody test, we examined on a fully automated enzyme immunoassay, is easy to assay in routine laboratory, and showed good analytical performance. And anti-CCP antibody test also showed higher diagnostic specificity than RF. So, anti-CCP antibody may be useful serologic marker for diagnosis and monitoring of RA, if performed concomitantly with RF assay.
Antibodies* ; Arthritis, Rheumatoid ; Diagnosis ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunoenzyme Techniques* ; Nephelometry and Turbidimetry ; Rheumatic Diseases ; Rheumatoid Factor ; Sensitivity and Specificity

At the summer workshop of the Korean Endocrine Society held in 2016, some examples of protein experiments were discussed in the session entitled “All about working with proteins.” In contrast to what the title suggested, it was unrealistic to comprehensively discuss all protein analytical methods. Therefore, the goal was to outline protein experimental techniques that are useful in research or in bench work. In conversations with clinicians, however, I have always felt that researchers who do not engage in bench science have different demands than those who do. Protein research tools that are useful in bench science may not be very useful or effective in the diagnostic field. In this paper, I provide a general summary of the protein analytical methods that are used in basic scientific research, and describe how they can be applied in the diagnostic field.
Chromatography ; Education ; Immunoenzyme Techniques ; Methods* ; Molecular Imaging

BACKGROUND: Human parvovirus B19 (B19) is associated with a variety of clinical infections. As B19 antigen is not available in large quantity, studies on cloning and recombinant protein expression have been performed. We cloned and expressed VP2 protein of B19 viruses to obtain B19 antigen in large quantity. METHODS: VP2 gene was amplified by PCR. pQE vector was constructed and transformed into E. coli for the expression of recombinant VP2 protein. Fusion protein was purified by Ni-NTA protein purification system. Finally, the purified antigen was tested for ELISA to detect IgG and IgM antibodies in patients. RESULTS: Recombinant VP2 protein, produced by transformed E. coli and expressed, was confirmed to be 58 kD protein by Western blot. Antibodies were detected in sera from all of eight patients, who had been suspicous of B19 infection clinically and confirmed to harbor B19 DNA by PCR, in assays using ELISA kit coated with recombinant VP2. CONCLUSIONS: VP2 protein, purified by recombinant protein expression in this study, can be used for enzyme immunoassay for the detection of serum antibodies and can be devoted to the future study on vaccine development.
Antibodies ; Blotting, Western ; Clone Cells* ; Cloning, Organism* ; DNA ; Enzyme-Linked Immunosorbent Assay ; Humans* ; Immunoenzyme Techniques ; Immunoglobulin G ; Immunoglobulin M ; Parvovirus B19, Human* ; Polymerase Chain Reaction

BACKGROUND: Clostridium difficile is one of the most important pathogens responsible for nosocomial diarrhea. The disease is mediated by two toxins, designated as A and B; therefore, identification of the toxins is important for diagnosis. However, culture or cytotoxin assay are not easily done because of tedious procedures. Instead, toxin A immunoassay is widely used. We evaluated two different enzyme immunoassays (EIA) for C. difficile toxin A and compared them with culture and PCR results. METHODS: A total of 65 stool specimens were examined for toxin A using enzyme linked fluorescent immunoassay (ELFA, VIDAS CD II, Bio-Merieux, France) and enzyme linked immunosolvent assay (ELISA, C.DIFFICILE TOX A II, TECHLAB, USA ) and were also cultured for C. difficile using cycloserine cefoxitine fructose agar. We amplified toxin A and B genes using primers NK9-NK 11 and NK104-NK105, respectively, in 23 C. difficile isolates. RESULTS: The concordance rate between ELFA and ELISA was 76.9%. The sensitivity and specificity of the ELFA and ELISA based on the culture and PCR results for toxin A gene were 84.6%/98.1% and 84.6%/67.3%. Positive and negative predictive values were 91%/96.2% in VIDAS and 78.0%/ 94.6% in TECHLAB. The positive rates of toxin B genes were 100%, 83.3% and 50% in toxin A positive, variant and negative strains, respectively. CONCLUSIONS: The sensitivities of the ELFA and ELISA for toxin A were the same, but specificity and positive predictive value of the ELFA were higher than those of the ELISA. PCR or EIA method detecting both toxin A and toxin B is strongly recommended, because the variant strains (toxin A negative and toxin B positive) of C. difficile may be more prevalent than were anticipated in Korea.
Agar ; Cefoxitin ; Clostridium difficile* ; Clostridium* ; Cycloserine ; Diagnosis ; Diarrhea ; Enzyme-Linked Immunosorbent Assay ; Fructose ; Genes, vif ; Immunoassay ; Immunoenzyme Techniques* ; Korea ; Polymerase Chain Reaction ; Sensitivity and Specificity