BACKGROUND: An accurate ABO blood type is essential for transfusion therapy. Genetic structures of ABO blood group and subgroup have been investigated and so far about 100 ABO alleles have been reported worldwide. This study was performed to investigate the molecular characteristics of B subgroups in the Korean population. METHODS: A total of 19 samples of B subgroups were collected from patients (n=11) and from blood donors (n=8) of Korean Red Cross blood centers; these samples had been typed serologically for the ABO blood group. Allele-specific polymerase chain reaction (PCR), direct sequencing of exon 6 and 7, and allele separation were performed for ABO gene analysis. RESULTS: The ABO PCR-RFLP genotyping results of 18 samples among the provisional 19 B subgroups were identical regardless of their phenotypes. Two new B alleles showing 255C>T base change and 547G>A base change were observed in B3 and A1B3 subgroups. CONCLUSIONS: Serologically unidentified B subgroups were unequivocally identified through molecular analyses of the ABO gene. And new ABO alleles observed only in the Korean B subgroups were recognized.
Alleles ; Asian Continental Ancestry Group ; Blood Donors ; Exons ; Genetic Structures ; Humans ; Phenotype ; Polymerase Chain Reaction ; Red Cross

BACKGROUND: We intended to confirm a decline in neutrophil function with aging and to devise a simple neutrophil function test protocol for use in a clinical setting. Reversely, the reliability of this protocol was to be verified by detectability of a decline in neutrophil function with aging. METHODS: Whole blood samples from young (thirties, N=32) and old (sixties, N=32) healthy subjects were incubated with the 7-aminoactinomycin D stained Escherichia coli. The mixture was stained by dihydrorhodamine 123 as an oxidative probe. Two kinds of fluorescence were measured by flow cytometry. RESULTS: Phagocytosis was declined with aging as indicated by a decrease in the percentage (form 28.2+/-9.5% to 21.9+/-10.9%, P<0.05) and the mean fluorescence intensity (MFI) ratio (from 1.67+/-0.27 to 1.51+/-0.27, P<0.05). Oxidative burst had a trend toward a decrease with aging, but the differences were not significant (percentage: from 35.3+/-13.2% to 32.1+/-14.1%, P=0.36; MFI ratio: from 5.26+/-3.23 to 5.08+/-3.55, P=0.84). CONCLUSIONS: The devised protocol in this study could detect a significant decline in neutrophil function with aging, and this protocol may be useful for the assessment of neutrophil function in a clinical setting.
Aging ; Escherichia coli ; Flow Cytometry ; Fluorescence ; Neutrophils* ; Phagocytosis ; Respiratory Burst

BACKGROUND: In the previous study, we reported a decrease of CD4-CD8- double negative T (DNT) cells with aging. In this study we investigated the precise changes of two subsets of DN T cells, alpha beta TCR DN T cells and gamma delta TCR DN T cells, with aging. METHODS: We analyzed the T cell subsets by 3 color flow cytometry in a healthy young group (age 26.9+/-2.5, N=15), a healthy old group (age 67.7+/-3.4, N=15) and an old group with aging-associated diseases. RESULTS: The percentage of total DN T cells, alpha beta TCR DN T cells and gamma delta TCR DN T cells in total T cells, respectively, was 7.4+/-4.9%, 3.9+/-2.9% and 3.5+/-2.5% in a healthy young group and 3.5+/- 2.2%, 1.8+/-1.6%, 1.6+/-1.5% in a healthy old group. Both subsets decreased with aging significantly both P<0.05). Between a healthy old group and an old group with aging-associated diseases, we could not detect the significant difference of the percentage of either subset. CONCLUSIONS: This study confirmed our previous report. Furthermore, we observed that both subsets of DN T cells decreased in detail. More studies are needed to clarify the association between these findings and the aging-associated diseases.
Aging* ; Flow Cytometry ; T-Lymphocyte Subsets ; T-Lymphocytes*

Members of the genus Hantavirus are the etiologic agents of hemorrhagic fever with renal syndrome (HFRS), the diagnosis of which is somewhat difficult because several diseases share similar early clinical presentations such as fever and petechia. In Korea, Hantaan virus and Seoul virus are the causative organisms of HFRS, and the infection caused by Seoul virus is milder than that caused by Hantaan virus. We report a 44-year-old woman, who visited our hospital due to general weakness, fever, myalgia, facial edema and diarrhea. She was diagnosed with HFRS caused by Seoul virus. The antibody against Hantaan virus was positive by an indirect immunofluorescent test and the discrimination between Hantaan and Seoul viruses was done by RT-PCR-RFLP (reverse transcriptionpolymerase chain reaction-restriction fragment length polymorphism) against viral S segment.
Adult ; Diagnosis ; Diarrhea ; Discrimination (Psychology) ; Edema ; Female ; Fever ; Hantaan virus ; Hantavirus ; Hemorrhagic Fever with Renal Syndrome* ; Humans ; Korea ; Myalgia ; Seoul virus*

Arcobacter butzleri, a gram-negative, slightly curved bacterium previously described as an aerotolerant Campylobacter, was isolated from the blood of a 53-year-old male patient with alcoholic cirrhosis. The isolate was identified by various phenotypic tests and 16S rRNA sequencing analysis. The patient was treated with amikacin and recovered uneventfully. To our knowledge, this is the first case of A. butzleri bacteremia reported in Korea.
Amikacin ; Arcobacter* ; Bacteremia* ; Campylobacter ; Humans ; Korea ; Liver Cirrhosis, Alcoholic ; Male ; Middle Aged

BACKGROUND: Clinical isolates of Escherichia coli were evaluated to determine the prevalence and genotypes of Ambler class A extended-spectrum beta -lactamases (ESBLs). METHODS: Clinical isolates of E. coli were collected from 12 hospitals from February through July, 2004. Antimicrobial susceptibility was tested by disk diffusion and agar dilution methods, and ESBLproduction was determined by double-disk synergy test. TEM, SHV, CTX-M, PER-1, VEB, IBC, GES, and TLA type ESBL genes were detected by PCR amplifications, and the PCR products were subjected to direct sequencing. RESULTS: The double-disk synergy test was positive in 90.9% (149 in 164) of the ceftazidime- or cefotaxime-resistant E. coli isolates. The most prevalent types of Ambler class A ESBLs in E. coliisolates were CTX-M-15 (n=53). CTX-M-14 (n=24), CTX-M-3 (n=9), CTX-M-9 (n=3), CTX-M-12 (n=3), SHV-2a (n=1), SHV-12 (n=5) and TEM-52 (n=3) were also found. CTX-M-12 ESBL had never been reported before in Korea. CONCLUSIONS: CTX-M type ESBL-producing E. coli isolates are spreading and CTX-M-12 is emerging in Korea.
Agar ; beta-Lactamases* ; Diffusion ; Escherichia coli* ; Genotype ; Korea ; Polymerase Chain Reaction ; Prevalence

BACKGROUND: Recently, the disk diffusion testing of fluconazole against Candida spp. has been attempted in order to provide a simple inexpensive method in the routine laboratory. We investigated the possibility and reliability of a fluconazole disk diffusion method using a Mueller-Hinton agar supplemented with glucose and methylene blue (GM-MH). METHODS: One hundred and seven isolates of Candida spp. (54 C. albicans, 21 C. glabrata, 20 C. tropicalis, 6 C. parapsilosis, 4 C. krusei, and 2 C. lusitaniae) were tested with the broth microdilution method of National Committee for Clinical Laboratory Standards (NCCLS) document M27-A2 and a disk diffusion test using GM-MH agar. RESULTS: The overall categorical agreement between the NCCLS method and the disk diffusion method was 89.8% for fluconazole, with 0.9% very major errors and 9.3% minor errors; no major errors were detected. CONCLUSIONS: These data suggest that the fluconazole disk diffusion test on GM-MH agar can be used as a routine screening procedure for susceptibility of Candida spp. in the clinical laboratory.
Agar* ; Candida* ; Diffusion* ; Fluconazole* ; Glucose ; Mass Screening ; Methylene Blue

BACKGROUND: The vanA gene cluster of vancomycin-resistant enterococci (VRE) is carried as a part of Tn1546-like elements. In this study we characterized the structure of Tn1546-like elements in Enterococcus. faecium isolated from patients in Korea. The isolates were also typed by pulsed-field gel electrophoresis (PFGE). METHODS: During 2000, 21 clinical isolates of vanA-containing E. faecium were collected from ten university hospitals in Korea. E. faecium BM4147 was used as a control. PFGE was performed on a CHEF-DR III apparatus. For structural analysis of Tn1546, the overlapping PCR amplification of internal regions of Tn1546 was performed. The purified PCR products were directly sequenced by using ABI Prism 3100 DNA SEQUENCER. RESULTS: All isolates were divided into 3 types according to the distribution of insertion sequences (IS elements) integrated Tn1546 elements. Type I and II were characterized by an IS1542 insertion in the orf2-vanR intergenic region and an IS1216V insertion in the vanX-vanY intergenic region. Type III represented two copies of IS1216V at the orf1 and in the vanX-vanY intergenic region as well as IS1542 in the orf2-vanR intergenic region. No isolates were identical to the prototype, which was identical to the predicted pattern for the published sequence of Tn1546. The PFGE results revealed that all strains except A13, C1, A2 and A9 were genetically unrelated. CONCLUSIONS: The distribution of IS in Tn1546-like elements of the Korean isolates is similar to that of the European VREs. Considering the results of PFGF and Tn1546 typing, the horizontal transfer of vanA resistance gene may be occurring among genetically diverse strains of E. faecium in Korea.
DNA ; DNA, Intergenic ; Electrophoresis, Gel, Pulsed-Field ; Enterococcus ; Hospitals, University ; Humans ; Korea ; Multigene Family ; Polymerase Chain Reaction

BACKGROUND: Homocysteine (Hcy) is known to increase the risk of cerebrovascular disease. I investigated this association in cerebral infarction (CI) and established reference intervals for serum total Hcy concentrations among individuals aged 40 or over in Korea. METHODS: I measured Hcy concentrations in the sera from 93 healthy controls (male 74, female 19) and 742 patients with CI (male 616, female 126) by a fluorescent polarization immuno assay technique using Axsym system (Abbott Laboratories, IL, USA). RESULTS: In CI group, the following parameters were significantly higher (P<0.001) than in normal control (NC): the serum Hcy level (13.4+/-6.6 vs. 10.3+/-2.6 micromol/L), age (67.1+/-9.0 vs. 62.6+/-9.9 years), C-reactive protein (10.2+/-22.2 vs. 5.5+/-6.4 mg/dL), and the prevalence of hypertension (69 vs. 33%) and diabetes mellitus (DM) (38 vs. 5%). The serum folate level in CI group was significantly lower than in NC group (7.5+/-4.1 vs. 8.8+/-5.0 ng/mL, P=0.037). The mean and upper reference limit of Hcy in male control (10.7 and 14.9 mol/L) were significantly higher than in female control (8.1 and 10.8 micromol/L). The risk of CI was higher in subjects with old age (> or =60 years), hypertension, DM, hyperhomocysteinemia, high creatinine, and in the highest Hcy quartile (> or =15.1 micromol/L) compared to the lowest Hcy quartile (<9.6 micromol/L) with the crude odds ratios of 2.1, 4.3, 10.5, 7.4, 3.0, and 6.6, respectively; in multivariate analysis, the risk of CI was independently associated with hypertension, DM, hyperhomocysteinemia and adjusted odds ratios were 3.6, 5.3, and 7.1, respectively. In CI group, Hcy exhibits negative correlations (P<0.001) with folate (r=-0.356) and vitamin B12 (r=-0.256). CONCLUSIONS: Hyperhomocysteinemia may represent an independent risk factor for cerebrovascular disease.
C-Reactive Protein ; Cerebral Infarction* ; Creatinine ; Diabetes Mellitus ; Female ; Folic Acid ; Homocysteine ; Humans ; Hyperhomocysteinemia* ; Hypertension ; Korea ; Male ; Multivariate Analysis ; Odds Ratio ; Prevalence ; Risk Factors* ; Vitamin B 12

BACKGROUND: C-reactive protein (CRP) concentrations are measured by two different assays depending on the clinical concern: the conventional CRP assay for estimating the extent of inflammation and the high sensitivity-CRP (hs-CRP) assay for assessing the risk of cardiovascular diseases. To integrate the conventional CRP test detecting acute phase response and the hs-CRP assay with a lower detection limit, we evaluated the performance characteristics of hs-CRP assay methods with a wide range of concentrations. METHODS: Immunonephelometric assay (BNII) and two turbidoimmunometric assays (TIA), the Hitachi 7600 with Daiichi reagent (Daiichi-Hitachi) and the Roche Cobas Integra 700 (Integra), were evaluated for the precision with 8 levels of pooled sera ranging from 0.3 to 120 mg/L and the agreement of TIA with the BNII assay using regression analysis and Bland-Altman analysis with 89 patient samples. RESULTS: The within-day coefficients of variation (CVs) of BNII were less than 10% in all levels of pooled sera. The CVs of Daiichi-Hitachi and Integra exceeded 10% in pooled sera below 1.0 mg/L and 0.5 mg/L, respectively. Both Daiichi-Hitachi and Integra appeared to be linear over the entire range of CRP concentrations used and were comparable with the results of BNII (Daiichi-Hitachi: y=0.98x+0.13, r=0.97, Integra; y=1.02x+0.22, r=0.99). However, at the concentrations over 100 mg/L TIA and BNII showed discrepant results. CONCLUSIONS: Both Daiichi-Hitachi and Integra showed a good precision over a wide range of CRP. However, the discrepant results found at very high concentrations require standardization among different assay methods or instruments.
C-Reactive Protein* ; Cardiovascular Diseases ; Humans ; Inflammation ; Limit of Detection