BACKGROUND: The serological detection of the surface antigen (HBsAg) of hepatitis B virus (HBV) is the basis of detection of HBV infections in blood donors and patients with hepatitis. The aim of this study was to compare the performance of HBsAg enzyme-linked immunosorbent assay (ELISA) and HBsAg chemiluminescence immunoassay used in Korea. METHODS: We compared seven assays: Architect i 2000 (Abbott Laboratories, USA), Elecsys 2010 immunoanalyzer (Roche Diagnostics, Germany), Advia Centaur (Bayer Healthcare, USA), Murex HBsAg version 3 (Abbott Laboratories, USA), Enzygnost HBsAg 5.0 (DADE Behring, Germany), LG HBsAg ELISA (LG, Korea), and Genedia HBsAg ELISA 3.0 (Greencross Medical Science, Korea). We evaluated the sensitivity of each assay by testing serially diluted WHO HBsAg reference material, two seroconversion panels, and recombinant HBsAg with three mutations in the 'a' determinant. RESULTS: The lowest HBsAg level detected by each assay using WHO reference material was variable from 0.05 (Murex and Advia) to 0.2 IU/mL. When testing 21 seroconversion panels, the total number of positive samples was 15 by Murex and 14 by Architect. Murex, LG, and Architect detected all of the 3 mutant samples tested. CONCLUSIONS: Analytical sensitivity and mutant detecting ability among HBsAg commercial assays were variable and not related to the analytical methods, but related to the manufacturer's reagents. We suggest that each laboratory should select an HBsAg assay based on analytical performance, test throughput, and the applicability of full automation.
Chemiluminescent Measurements/*methods ; Enzyme-Linked Immunosorbent Assay/*methods ; Hepatitis B Surface Antigens/*blood ; Humans ; Reagent Kits, Diagnostic ; Reference Values ; Sensitivity and Specificity

BACKGROUND: An increased level of beta2-microglobulin (beta2M) is seen in diseases such as lymphoproliferative diseases, renal diseases, solid tumors, liver diseases, certain viral infection, or chronic inflammatory diseases, etc. In this study, we evaluated a quantitative beta2M assay for precision and linearity using an automated turbidimetric immunoassay (TIA) by Hitachi 7600-110 (Hitachi High Technologies Co., Japan). The TIA of beta2M was compared with a chemiluminescent immunoassay (CLIA) by Immulite 2000 (Diagnostic Products Corporation, USA). METHODS: Two TIAs, the Hitachi 7600-110 with Roche reagent (Roche-TIA) and the Hitachi 7600- 110 with HBI reagent (HBI-TIA), were evaluated for within-run precision, within-day precision, betweendays precision, and linearity. With 68 serum samples, two TIAs were compared with Immulite 2000 using DPC reagent (DPC-CLIA). These data were analyzed by Passing-Bablok analysis and Bland- Altman analysis. RESULTS: The coefficients of variation (CVs) of within-run precision, within-day precision, and between- days precision were less than 7% in all groups. The linearity tests of the two TIAs were maintained well (Roche-TIA: R2=0.9952; HBI-TIA: R2=0.9946). The comparison study indicated good correlations (Roche-TIA/DPC-CLIA: r=0.9738, y=0.9625x-0.0375; HBI-TIA/DPC-CLIA: r=0.9725, y= 1.1000x-0.3100). In Bland-Altman analysis, less than 2SD differences were observed between the two groups. CONCLUSIONS: Both Roche-TIA and HBI-TIA showed a good precision and excellent correlations with DPC-CLIA; therefore, TIA could be used in the routine laboratory to determine a quantitative analysis of beta2M.
Aged ; Chemiluminescent Measurements ; Enzyme Multiplied Immunoassay Technique ; Female ; Humans ; Immunoassay/*methods ; Male ; Middle Aged ; Nephelometry and Turbidimetry ; Reagent Kits, Diagnostic ; Regression Analysis ; Sensitivity and Specificity ; beta 2-Microglobulin/*analysis

The luminous intensity of dark variant (S1) separated from photobacterium phosphoreum (A2) was 1/10,000 less than that of wild-type. Ethidium bromide (EB) (0.6 mg/L), Mytomycin C (MC, 0.05 mg/L), 2-amino fluorene (2-AF, 1.0 mg/L) all could strongly induce reversion mutation for S1 within 24 h and increase reversion ratio significantly. The results of experiments indicated that these revertants had stable genetic characteristic and the mutation may take place at gene levels. The mutagenesis to S1 caused by EB, MC and 2-AF was detected and it may be used as a new rapid, simple and sensitive method for gene toxicant monitoring.
*Chemiluminescent Measurements ; Ethidium/pharmacology ; Ethidium/toxicity ; Luciferases/biosynthesis ; Mitomycins/pharmacology ; Mitomycins/toxicity ; Mutagens ; Mutation/*drug effects ; Photobacterium/*genetics ; Toxicology/methods ; Transcription, Genetic/drug effects ; Variation (Genetics)

BACKGROUND: Anti-double stranded DNA antibody (anti-dsDNA) test is useful for the diagnosis and monitoring of systemic lupus erythematosus (SLE). Although several methods are available, none of them is completely satisfactory and differences among them have been reported. We evaluated the diagnostic performance of 6 commercial kits for anti-dsDNA detection. METHODS: A total of 142 sera (SLE [N=74], other systemic rheumatic diseases [N=50], other diseases [N=18]) were tested by 6 different assay kits using different antigenic sources of DNA: Crithidia luciliae immunofluorescence test (CLIFT), salmon testes (immunoblot, IB), human (ELISA I), salmon testes with nucleosome linker (ELISA II), plasmid (ELISA III), and synthetic oligonucleotides (chemiluminescence immunoassay, CLIA). RESULTS: With manufacturers' cut-off values, 6 test kits showed sensitivities of 55.4-91.9%. ELISA I had a greater sensitivity than the other five assays (P<0.001). The specificities of ELISA II, ELISA III, CLIA, and CLIFT were higher than those of ELISA I and IB (P<0.05). In ROC curve analysis, 3 ELISA kits and CLIA showed AUC values of 0.845-0.893, and revealed no significant differences among them (P>0.05). With cut-off values set at 95% of specificity, ELISA II had a higher sensitivity than ELISA III (63.5% vs. 41.9%, P<0.05). IB had poor concordance rates with other assays (42.0-65.0%). Pearson correlation coefficients among 4 quantitative assays were 0.667-0.798. CONCLUSIONS: Six different assays showed various performances depending on the methods and cut-off values used. Except IB, the other five assays can be used for the detection of anti-dsDNA.
Adult ; Antibodies, Antinuclear/*analysis ; Area Under Curve ; Chemiluminescent Measurements/*methods ; DNA/*immunology ; Enzyme-Linked Immunosorbent Assay/*methods ; Female ; Fluorescent Antibody Technique/*methods ; Humans ; Immunoblotting/*methods ; Lupus Erythematosus, Systemic/diagnosis ; Male ; Middle Aged ; ROC Curve ; Reagent Kits, Diagnostic ; Sensitivity and Specificity

BACKGROUND: Hepatitis C virus (HCV) core antigen (Ag) levels are known to be well correlating with HCV RNA levels, and may be used as an alternative marker of HCV replication for monitoring the response to HCV treatment. However, the low sensitivity of HCV core Ag assay has been an obstacle for clinical use. In this study, recently developed ARCHITECT HCV Ag assay (Abbott Laboratories, USA) was evaluated for analytical performance and clinical usefulness. METHODS: A total of 109 sera from HCV infected patients including various genotypes of HCV (1b, 2, 2a/2c, 2b, and 3a) and 20 sera from healthy donors were used for evaluating the sensitivity, precision, and linearity of the HCV core Ag assay. The cross reactivity with HIV, hepatitis B virus and myeloma proteins (N=5, each) and correlation with HCV RNA PCR assay were also evaluated. RESULTS: The sensitivity of the HCV core Ag assay was 97.2% (106/109) and there were no false positive results and cross reactivity. The within-run, between-run and between-day CVs were 3.0%, 2.5% and 3.0%, respectively. The levels of HCV core antigen showed a good correlation with those of HCV RNA quantification (r=0.940). The HCV Ag assay showed an excellent linearity in the range from 0.63 to 17,114 fmol/L (r=0.999). CONCLUSIONS: The ARCHITECT HCV Ag assay was good in sensitivity, precision, and linearity and its results well correlated with HCV RNA levels. This assay could be used as a good marker of viral replication for monitoring the therapy response in chronically HCV infected patients.
Chemiluminescent Measurements/*methods ; Cross Reactions ; Genotype ; Hepacivirus/genetics/*immunology ; Hepatitis Antigens/*blood ; Humans ; Polymerase Chain Reaction/*methods ; RNA, Viral/blood ; Reagent Kits, Diagnostic ; Sensitivity and Specificity ; Viral Core Proteins/*blood

BACKGROUND: The Lewis histo-blood group system consists of 2 major antigens-Lea and Leb-and a sialyl Lewis antigen-carbohydrate antigen (CA) 19-9. We investigated the distribution of Lewis genotypes and evaluated the relationship between the Lewis/Secretor genotypes and the serum level of CA 19-9 in a Korean population to identify whether the serum CA 19-9 levels are influenced by the Lewis/Secretor genotypes. METHODS: The study included 242 individuals who had no malignancies. Lewis genotyping was performed for the 59T>G, 508G>A and 1067T>A polymorphic sites. The Secretor genotype was determined through analysis of the 357C>T and 385A>T polymorphic sites and the fusion gene. Serum CA 19-9 level was analyzed using an electrochemiluminescence immunoassay. RESULTS: Individuals carrying the 3 common genotypes-Le/Le, Le/le(59,508), and Le/le(59,1067)-accounted for 95% of the study population. In the Korean population, the allelic frequencies of Le, Le(59), le(59,508), and le(59,1067) were 0.731, 0.010, 0.223, and 0.035, respectively. We found a significant difference in serum CA 19-9 concentrations among the 9 Lewis/Secretor genotype groups (P<0.001). The serum CA 19-9 levels in subjects with genotype groups 1 and 2 (Le/- and se/se) were higher than those with genotype groups 3-6 (Le/- and Se/-; 15.63 vs 6.64 kU/L, P<0.001). CONCLUSIONS: Le/Le, Le/le(59,508), and Le/le(59,1067) are frequent Lewis genotypes in Koreans. Because serum CA 19-9 levels are significantly influenced by the Lewis/Secretor genotypes, caution is suggested when interpreting the serum CA 19-9 levels.
Adult ; Aged ; Alleles ; Asian Continental Ancestry Group/*genetics ; CA-19-9 Antigen/*blood ; Chemiluminescent Measurements/methods ; Female ; Gene Frequency ; Genotype ; Humans ; Immunoassay/methods ; Lewis Blood-Group System/*genetics ; Male ; Middle Aged ; Phenotype ; Polymorphism, Genetic ; Republic of Korea

Autoantibodies against thyroxin (T4AA) and triiodothyronine (T3AA) are present in dogs with autoimmune thyroiditis and have been reported to interfere with immunoassays. The objectives of this study were to determine the frequency of autoantibodies and to determine whether interference occurs by T4AA, using a non-immunological method (high performance liquid chromatography, HPLC) for thyroxin (T4) measurement. Based on clinical symptoms, T4 and thyroid stimulating hormone (TSH) concentration, 1,339 dogs were divided into six groups: Group 1: hypothyroid (n = 149); Group 2: subclinical thyroiditis (n = 110); Group 3: suspicious for non thyroidal illness (n = 691); Group 4: biochemical euthyroid (n = 138); Group 5: hypothyroid dogs under substitution therapy (n = 141); Group 6: healthy dogs (n = 110). The incidence of T4AA and T3AA, determined using radiometric assay, was low (0.5% and 3.8%) and higher in hypothyroid dogs compared to dogs suspicious for hypothyroidism (Group 2-4) (p<0.05). T4AA was not detected in dogs with normal T4 and elevated TSH. T4 concentrations of T4AA positive samples determined using HPLC were comparable to results obtained by chemiluminescence immunoassay. These findings indicate that the probability of interference of T4AA leading to falsely elevated T4 concentration in the T4 assay seems to be low.
Animals ; Autoantibodies/*immunology ; Chemiluminescent Measurements/methods/*veterinary ; Chromatography, High Pressure Liquid/veterinary ; Dog Diseases/*diagnosis/*immunology ; Dogs ; Immunoassay/methods/*veterinary ; Thyroid Hormones/*immunology ; Thyroiditis, Autoimmune/diagnosis/immunology/*veterinary ; Thyroxine/*blood

Many allergists are currently focusing on the development of new diagnostic tools, and are attempting to improve both the sensitivity and specificity. A multiple allergen simultaneous test-chemiluminescent assay (MAST-CLA) is one of the most popular diagnostic tools used in the Republic of Korea. However, there remains controversy among allergists with regard to the cut-off point for a positive result. The present study was conducted in order to determine the validity of MAST-CLA as compared with that of the skin prick test, with particular emphasis on arthropod allergens, on the basis of percentage agreement rates and k-values, and also to suggest the optimal positive cutoff points using receiver operating characteristic (ROC) curves. The study was conducted with 97 subjects (54 men, 43 women). Optimal individual cut-off points were calculated as follows; class II for Dermatophagoides farinae, class I for Dermatophagoides pteronyssinus, and trace for a cockroach mix. These findings suggest that attempting to apply optimal individual cut-off points will be a good way of improving diagnostic tests, particularly MAST-CLA.
Adult ; Allergens/*immunology ; Animals ; Antigens, Dermatophagoides/*immunology ; Chemiluminescent Measurements/*methods/standards ; Cockroaches/chemistry ; Dermatophagoides farinae/chemistry ; Dermatophagoides pteronyssinus/chemistry ; Female ; Humans ; Hypersensitivity/*diagnosis/immunology ; Insect Proteins/*immunology ; Male ; *ROC Curve ; Skin Tests/methods