Among the fifty-three clinical isolates of Haemophilus influenzae, nineteen isolates including eight isolates of each biotype I-VIII, six of serotype b (Hib) strains and five of nontypeable strains were characterized by SDS-PAGE about outer membrane protein (OMP), restriction enzyme analysis (REA) and rRNA gene restriction pattems. OMP patterns showed to common band patterns in each H. influenzae isolate. Based on the two major proteins, 31KDa-38KDa, isolated strains were classified into 7 subtypes. In the OMP patterns about biotype and serotype, the specific pattern of each biotype was not distinguishable, but all of the serotype b strains were shown identical unique pattern, therefore it made distinctive difference with nontypeable strains. The digested genomic DNAs with EcoRI were identical result with rRNA gene restriction. It was more subdivided into 10 ribotypes. The most common ribotype I and serotype 1 accounted for 6 strains (31.6%) and 7 strains (36.8%) of the 19 clinical isolates, respectively. Hib isolates that were both OMP subtype 1 and ribotype I accounted for 2 strains (10.5%). In the epidemiologically unrelated strains, the putative association between the subtypes could not be confirmed. According to these results, the three methods were discriminatory and appropriate techniques for epidemiological studies of H. influenzae.
DNA ; Electrophoresis, Polyacrylamide Gel* ; Genes, rRNA* ; Haemophilus influenzae type b ; Haemophilus influenzae* ; Haemophilus* ; Influenza, Human ; Membrane Proteins ; Restriction Mapping* ; Ribotyping

No abstract available.
Antibody Formation* ; Borrelia burgdorferi* ; Borrelia* ; Retrospective Studies*

No abstract available.
Haemophilus influenzae* ; Haemophilus*

PURPOSE: In this study we tried to look at the spreading, duration of colonization, and acquisition of new streptococci which were obtained in one geographical area, as well as the bacteriologic and molecular epidemiology of normal school children carrying group A streptococci and their clonal relationship through the combined application of the serotype of T antigen and Pulsed Field Gel Electrophoresis(PFGE). METHODS: A total of 88 strains of group A streptococci were isolated from 396 normal school children. All isolates were classified in groups by Streptex and serotyped by T. agglutination. Restriction enzyme digestion of DNA was taken using Sma I. DNA fragments were separated by PFGE. RESULTS: A total of 33 strains were allocated their epidemiologic characteristics. Four out of 33 strains were not restricted by enzyme(Sma I). Twenty nine strains out of 33 strains showed 12 subtypes with 8-12 fragments between 40kbp and 500kbp of DNA fragments on PFGE. Eight strains of NT and T6 war same fragment patterns on PFGE, respectively. Three strains out of 4 strains of T8/25 were not restricted and the other one showed different, unique patterns. One strain out of 8 stains of T12 was not restricted, and the others were classified as 5 different subtypes. Two strains of Tl were different patterns from each other, and 2 strains of T4 showed the samefragment pattern CONCLUSION: T serotypes with PFGE will be useful as a screening and molecular epidemiologic method in a country where anti-M antisera is not available, after recognizing the advantages and disadvantages of M and T serotyping.
Agglutination ; Antigens, Viral, Tumor ; Child* ; Colon ; Coloring Agents ; Digestion ; DNA ; Epidemiologic Methods ; Epidemiologic Studies* ; Humans ; Immune Sera ; Mass Screening ; Molecular Epidemiology ; Serotyping* ; Streptococcus pyogenes* ; Streptococcus*

PURPOSE: The identification of antigenic specificity of Streptococcus pyogenes using T serotyping is important to understand biologic characteristics of microorganisrns. We would like to disover the association of the occurrence of predominant T type, with possible outbreak of erythromycin resistant Streptococcus pyogenes in this country, which has been documented since the late 1990s. METHODS: Throat swab cultures were taken from a total of 1,294 normal school children(Subject A) in two different geographical areas. A total of 92 strains(Subject B) were obtained from the patients with group A streptococcal infections from Jan. 1998 to Dec. 1998. All strains were serotyped with T protein antisera. RESULTS: The distribution of T12 in Uljin increased from 4.2%(1996) to 45.7%(1998). T4 increased from 6.3% to 20.0%. Thirty-eight out of 92 strains were resistant to erythromycin. Twenty-seven out of 41 strains(T12) were multidrug resistant to erythromycin, clindarnycin, and tetracycline. CONCLUSION: We can see the sudden increase in T12 strains, one of the strains that are resistant to erythromycin in 1998, compared with previous years. T protein serotyping could be epidemiologically useful as a screening methods for detecting erythromycin resistant group A streptococci in hospitals where the routine antibiotic sensitivity test dose not examin for streptococci.
Epidemiology ; Epitopes ; Erythromycin* ; Humans ; Immune Sera ; Mass Screening ; Pharynx ; Population Characteristics ; Serotyping* ; Streptococcal Infections ; Streptococcus pyogenes ; Tetracycline

Ninety two strains of Streptococcus pyogenes were isolated from patients with pharyngitis, scarlet fever, skin infection, and invasive streptococcal infections in Seoul, Korea from January to December, 1998. All isolates were epidemiologically characterized by T protein serotype, and serum opacity factor (OF) detection to phenotypes. To analyze the genetic relationship, fifty two isolates including 32 erythromycin-clindamycin (Em-Cm) resistant strains, 20 antimicrobial susceptible strains were attempted to the pulsed-field gel electrophoresis (PFGE). T protein serotype showed 16 kinds in distribution including T12 and T4. Among the total isolates, 40 strains (43.5%) belonged to the T12 serotype and twenty strains (21.7%) to T4 serotype. On the other hand, when infection aspect of S. pyogenes isolates were analysed by T serotype distribution, T12 type was predominant for pharyngitidis which contributed to 21 strains (53%) and for skin infection isolates which contributed to 11 strains (28%), respectively. In case of T4 type, it was the most predominant pharyngitidis isolates which contributed to 8 strains (40%). In T serotype distribution of Em-Cm resistant strains, 27 strains (84%) of the thirty two showed T12 serotype. In minimum inhibitory concentration (MIC) values of Em-Cm resistance isolates, thirty two isolates showed resistant to erythromycin 27 strains (84%), had high MIC of >128 mug/ml. And also to clindamycin, twenty two strains (69%) had high MIC of >128 mug/ml. When OF detection of Em-Cm resistance of S. pyogenes isolates were analyzed by T serotype distribution, T12 serotype isolates revealed that all of the isolates except one strain were OF negative. In PFGE profile analysis to Em-Cm resistance isolates, of the twenty seven, Em-Cm resistance of T12 serotype isolates, 26 strains showed identical PFGE profile and all of these isolates revealed that OF negative. Eighty four percent of Em-Cm resistance S. pyogenes isolates had identical phenotype and PFGE profile. These results strongly suggested that the Em-Cm resistant S. pyogenes isolates from Seoul area showed close genetic correlation and PFGE could be available tool for molecular epidemiology.
Clindamycin ; Electrophoresis, Gel, Pulsed-Field* ; Erythromycin ; Hand ; Humans ; Korea* ; Microbial Sensitivity Tests ; Molecular Epidemiology ; Pharyngitis ; Phenotype ; Scarlet Fever ; Seoul ; Skin ; Streptococcal Infections ; Streptococcus pyogenes* ; Streptococcus*

A total of 152 strains of Streptococcus pyogenes were isolated from patients with pharyngitis, scarlet fever, skin infection, or invasive streptococcal infections in Seoul, Korea from January 1988 to December 1999. All isolates were epidemiologically characterized to decide phenotypes by T protein serotype and serum opacity factor (OF) detection. Genetic diversity of the isolates were analyzed by emm genotyping and pulsed-field gel electrophoresis (PFGE). T protein serotype showed 17 kinds in distribution and T12 (40.1% of study strains), T4 (19.1%), and T1 (7.9%) were the prevalent ones. When sources of S. pyogenes isolates were analyzed by T serotype distribution, T12 type was predominant in pharyngitis and skin infection isolates which contributed to 30 strains (49.2%) and 11 strains (18.0%), respectively. When T serotype of S. pyogenes isolates were analyzed by emm genotype distribution, of the 61 isolates of T12 type, 48 strains (78.7%) belonged to the emm type 12 (M12) and of the 29 isolates of T4 type, 27 strains (93.1%) belonged to the emm genotype 4 (M4). PFGE of genomic DNA of different emm genotype (emm12, emm4 and emm1) showed distinctive patterns. When the DNA of same emm gene type isolates were analyzed genetic relatedness by PFGE pattern, emm4, emm1, and emm12 types showed over 90%, 75%, and 70% of genetic similarity, respectively. Therefore, it was suggested that these emm genotype isolates were closely related genetically whereas among the isolates of other emm genotypes showed less than 30% of genetic similarity. Show genotypes are more diverse in comparison with phenotypes. In even epidemiologically unrealated isolates, genetic subtypes appeared correlated. The phenotypic and genotypic analysis used in the study were discriminative and appropriate for epidemiological study of S. pyogenes.
DNA ; Electrophoresis, Gel, Pulsed-Field ; Epidemiologic Studies ; Genetic Variation ; Genotype ; Humans ; Korea* ; Pharyngitis ; Phenotype ; Scarlet Fever ; Seoul* ; Skin ; Streptococcal Infections ; Streptococcus pyogenes* ; Streptococcus*

Species identification is an important item to characterize unidentified bacterial pathogens in developing and managing bacterial resources. In this study, unidentified pathogens based on the results of an automated identification system were identified using matrix assisted laser desorption ionization-time of flight mass spectrometry (MALD-TOF MS) and 16S rRNA gene analysis for development of national resources in the National Culture Collection for Pathogens (NCCP) in Korea. A total of 437 unidentified strains from branch banks of the NCCP were collected, and 16S rRNA and dnaJ gene sequencing, as well as MALDI-TOF MS analysis were performed to identify bacterial species. The mass spectra extracted were analyzed. Twelve strains exhibiting less than 98.65% similarity in 16S rRNA gene were selected as the primary candidates for novel species, and 21 strains exhibiting 98.65~99.0% similarity in 16S rRNA gene were selected as possible candidates for novel species. Among them, strain 32, belonging to Dermabacter sp., was finally selected as a possible strain representing a novel species and 14 unidentified bacterial strains using automated phenotypic identification system were newly registered at NCCP. The present study showed that unidentified pathogens using the automated phenotypic identification system were efficiently identified using the combination of MALDI-TOF MS and 16S rRNA gene analysis, and developed to the national resources in NCCP.
Genes, rRNA* ; Korea ; Mass Spectrometry

BACKGROUND: In this study, we compared various methods of taxonomic identification of Bacillus strains: biochemical methods, 16S rRNA gene sequencing, and matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS). We also developed a pathogen- isolate resource database, thus increasing the identification rate when using MALDI-TOF MS. METHODS: Thirty Bacillus strains were obtained from the NCCP (National Culture Collection for Pathogens) and were identified using the VITEK 2 system (bio-Mérieux, France), API kit (bioMérieux, France), 16S rRNA gene sequencing, and MALDI-TOF MS. The pathogenicity of Bacillus cereus was confirmed through the identification of virulent genes using a multiplex PCR, and both protein extraction for protein profiling in MALDI-TOF MS and repetitive-sequence fingerprinting were performed. RESULTS: The identification rates at the species level were 40%, 80%, and 76.3% for the VITEK 2 system (bioMérieux), 16S rRNA gene sequencing, and MALDI-TOF MS, respectively. When the major spectrum-profiling dendrogram was compared with the phylogenetic tree, which was constructed based on the 16S rRNA gene sequences and rep-PCR fingerprinting, the classifications were confirmed to be effective. CONCLUSION: Identification of Bacillus strains using MALDI-TOF MS was more effective than that using the VITEK 2 system (bioMérieux), but was similar to that using 16S rRNA gene sequencing. Continual addition to a proteome-based database can result in increased identification rates for MALDI-TOF MS.
Bacillus cereus ; Bacillus* ; Classification ; Dermatoglyphics ; Genes, rRNA ; Mass Spectrometry* ; Multiplex Polymerase Chain Reaction ; Trees ; Virulence

BACKGROUND: The zoonotic disease Q fever is caused by Coxiella burnetii and usually affects high-risk human populations. We conducted a serological survey of dairy cattle farmers in Korea to determine seroreactivity and identify risk factors for C. burnetii infection. METHODS: This cross-sectional study included 1,824 of 7,219 dairy cattle farms (25.3%) in the study region. The selected dairy cattle farmers visited the nearest public health centers or branches with completed questionnaires. Serum samples from the farmers were tested using an indirect immunofluorescence assay to detect phase II C. burnetii immunoglobulin (Ig) G or M antibodies. RESULTS: A total of 1,222 dairy cattle farmers from 784 dairy cattle farms (43.0%) participated in this study, and 11.0% (134/1,222) exhibited seroreactivity, defined as a phase II antigen IgG or IgM titer ≥ 1:16. In the multivariate analysis, male sex, residence in Gyeonggi Province, a larger herd size, and ocular/oral contact with birth products during calf delivery were significantly associated with a higher risk of C. burnetii infection. Furthermore, the risk was significantly lower among farmers who always wore protective gloves while cleaning cattle excretion, compared to those who sometimes or rarely wore protective gloves. CONCLUSION: Dairy cattle farmers should exercise caution by avoiding ocular/oral contact with birth products during calf delivery and by using protective equipment (including gloves).
Agriculture ; Animals ; Antibodies ; Cattle* ; Coxiella burnetii* ; Coxiella* ; Cross-Sectional Studies ; Farmers* ; Fluorescent Antibody Technique, Indirect ; Gloves, Protective ; Gyeonggi-do ; Humans ; Immunoglobulin G ; Immunoglobulin M ; Immunoglobulins ; Korea* ; Male ; Multivariate Analysis ; Parturition ; Public Health ; Q Fever* ; Risk Factors* ; Serologic Tests ; Zoonoses