No abstract available.
Escherichia coli O157* ; Escherichia coli* ; Escherichia* ; Polymerase Chain Reaction* ; Virulence Factors* ; Virulence*

Six strains of Vibrio parahemolyticus isolated from diarrheal patients and the 12 strains from sea water were serotyped and analyzed for biochemical characteristics, antibiotics sensitivity and detection of toxR, gyrB, tdh, and trh genes. Arbitrarily-primed polymerase chain reaction method were performed on the 6 strains from patients and the following results were obtained. 1. The Vibrio parahemolyticus isolated from patients were belonged to 5 different serotypes: 04:K8, 04:KUT, 06: K18, 010:K71 and 03:K6, but those isolated from sea water were belonged to 5 different serotypes: O1:KUT, 02:KUT, 03:K45, 04:K37 and OUT:KUT. All strains explained have different serotypes depending on the different source, 2. Three serotype (04:K8, 04:KUT, 06:K18) isolated from patients were positive for the urease hydrolysis, whereas only one strain of serotype O1:KUT isolated from sea water was positive to the same. Furthermore, the serotype 06:K18 (1 strain) was positive for the fermentation of dulcito1. Both toxR and gyrB genes were detected from all strains isolated. 3. As for control the 2 strains of serotype 03:K6 and 6 strains isolated from patients, serotype 03:K6 were resistant to oxacillin, penicillin, vancomycin. All strains were sensitive to chloramphenicol and tetracycline yet the antibiogram type showed 6 groups from I to VI. 4. DNA probe hybridization method was used to detect genes. The trh1 was detected both from serotype 04:KUT and 06:K18 isolated from patients and the trh2 was also detected from one strain from each 010:K71 and O1:KUT isolated from patients and sea-water respectively, The tdh gene only was detected from two strains of 03:K6 isolated from patients of 1998. The tdh, trh 1 and trh2 were not detected from 7 strains out of 12 strains isolated from sea water whereas the production titer of TDH isolated from patients showed from 2048 times to 4096 times. 5. Four strains of the serotype 03:K6 isolated from Korea, India and Japan as well as 3 strains from Korean patients were tested by AP-PCR to classify serotypes. As for its result the amplicon showed the same in the 4 strains of the serotype 03:K6 whereas the four strains of different serotype from patients are so difference as to explain no inter- relations at all. The result explains that the serotype 03:K6 is the same genes regardless from where it is isolated.
Anti-Bacterial Agents ; Chloramphenicol ; DNA ; Fermentation ; Humans ; Hydrolysis ; India ; Japan ; Korea ; Microbial Sensitivity Tests ; Oxacillin ; Penicillins ; Polymerase Chain Reaction ; Seawater* ; Tetracycline ; Urease ; Vancomycin ; Vibrio parahaemolyticus* ; Vibrio* ; Virulence Factors* ; Virulence*

No abstract available.
Aeromonas*

Fecal isolates of Escherichia coli which were collected from diarrheal patients and HUS patient in Pusan National University Hospital between 1990 and 1996, were serotyped and analyzed for plasmid DNA profile, biotype, HEp-2 cell adherence ability, reactivity to eae probe and for production of verotoxins (VT). In order to ease the diagnosis of EHEC infection, a LPS- based solid phase enzyme linked immunosorbent assay was utilized to detect serological diagnosis of EHEC infection. The following results were obtained. Among 150 EPEC isolates and HUS patient's stool, 7 EHEC were found. The 7 EHEC belonged to 5 different serotypes 0157:H7, 0143:H-, 0166:H-, 0128:H2, 026:H-, and 0111:H 21 previously associated with human haemorrhagic colitis (HC) and haemolytic uraemic syndrome (HUS). Biochemical cheracteristic analysis indicated 7 strains were biotype 1 and was found to have siderophilins suggesting their advantagous growth in vivo. For plasmid profiles, all strains had 60 MDa plasmid and several smaller sizes of plasmids. Three strains of Escherichia coli serotype 0157:H7, 0128:H2, and 026:H- showed one pattern of adherence in the HEp-2 cell assay namely, localized adherence and were positive for eae probe when tested by colony blot hybridization assay. PCR using specific primers for VT1, VT2 was tested, and all 7 strains carried VT1 gene only. PCR products of 130-bp (VT1) and 346-bp (VT2) were successfully amplified simultaneously in a single reaction. The multiplex PCR method can be used to specifically identify EHEC. The serum obtained from HUS patient of enterohemorrhagic E. coli were analyzed for rises in titer of intibody to somatic 02, 026, 0111, 0128, 0143, 0145, 0157, and 0165. Although response to the somatic 0 correlated significantly with response to the 026 rises of antibody titer to somatic 0 in acute stage of disease and anti-VT had not so many relation to that of VT. These results suggest that ELISA can be used to detect somatic 0 in serum and it is a useful method to diagnose the infection caused by EHEC rapidly.
Antibodies* ; Busan ; Colitis ; Diagnosis* ; DNA ; Enterohemorrhagic Escherichia coli* ; Enteropathogenic Escherichia coli ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; Humans ; Multiplex Polymerase Chain Reaction ; Plasmids ; Polymerase Chain Reaction* ; Shiga Toxins ; Transferrin

Sixteen strains of LT-producing enterotoxigenic E. coli 0128 which were isolated from diarrheal patient's stool in Pusan University Hospital, were serotyped and analyzed for plasmid DNA profile, MRHA of human blood cells, and also tested for possession of LT, ST, aggA, EAST1 genes by the PCR method and analyzed the RAPD pattern. Screening sensitivity for ETEC by salting out test was 87.5%. These data suggest that hydrophobicity test using salting out is rapid, inexpensive, and simple screening test for ETEC. CFAs were identified in 87.5% of strains; 43.75% the strains harbored CFA/I, 43.75% CFA/II, and 12.5% expressed none of these CFAs. For plasmid profiles, 12 strains had 60 MDa plasmid and several smaller plasmids. The strains showed 5 types of plasmid profiles. By PCR, LT gene but not ST gene was detected from all of the 16 strains EAST1 gene was detected from 14 strains. Ten strains could be differentiated to 3 patterns by chromosomal DNA fingerprint. The chromosomal DNA fingerprinting is considered very useful for the epidemiological study.
Blood Cells ; Busan ; DNA ; DNA Fingerprinting ; Enterotoxigenic Escherichia coli* ; Humans ; Hydrophobic and Hydrophilic Interactions ; Mass Screening ; Plasmids ; Polymerase Chain Reaction ; Virulence Factors* ; Virulence*

Cholera enterotoxin (CT) is a major virulence determinant of Vibrio cholerae 01. CI' is known to be the major virulence factor of Vibrio cholerae 01 and in accordance with the recent report showing which V. cholerae non-01 has ctx gene, we performed the molecular genetic study for the detection of ctx gene related to the production of CT at the subject Vibrio spp. except for V. cholerae non-01 and V. cholerae non-01 stock cultured in the laboratory of microbiology, College of Medicine, Pusan National University and the Vibrio spp. isolated from the marine products of Pusan General Fish Market and the sea water, and then its results are as follows: 1. PCR for the detection of ctx gene at the subject of V. cholerae 01:61H-151 having the ctx gene of which the denaturation is 1 rninute at 95'C, annealing to 1min, 30 sec at 60'C, the extension to be 1min. 30 sec at 72'C and 30 or 40 cycles. ctx gene was detected from 4 strains of V. cholera non-01 derived from the environment isolates. 2. Adjusting the quantity of chromosomal DNA used as template DNA to be from 0.1 pg to 1 ng, in order to know the PCR conditions for the effective search of ctx gene, and the detection limit of the system was 10 pg of chromosomal DNA. 3. The broth culture was used for template DNA, ctx gene of 302 bp was detected from 4 V. cholerae non-01, as in the case of chromosomal DNA, and the cell number was possible to be detected to 3 * 10.4. We attempted the confirmation of ctx gene through Southern blot hybridization, labeling with P and then it was confirmed only from 4 V. cholerae non-01 as like PCR results. 5. As the result of the sensitivity of PCR and Southern blot hybridization, it was shown to be possible which 10 pg was detected in case of chromosomal DNA and in case of cultured broth, the cell number was detected until 10 at PCR and Southern blot hybridization, and thus it was examed which its sensitivity was same.
Blotting, Southern ; Busan ; Cell Count ; Cholera Toxin* ; Cholera* ; DNA* ; Enterotoxins ; Limit of Detection ; Molecular Biology ; Operon* ; Polymerase Chain Reaction* ; Seawater ; Vibrio cholerae* ; Vibrio* ; Virulence

Eighty-two strains of Escherichia coli isolated from urine specimens in Pusan University Hospital, were serotyped and analyzed for plasmid DNA profiles, PFGE profiles, MRHA of human blood cells, HEp-2 cell adherence ability and reactivity to bfpA, LT, STh and STp DNA probes. The following results were obtained. Fifty-three of the eighty-two strains belonged to thirteen different 0 serotypes, twenty-nine strains could not be typed with the antisera used. Thirty strains (43.9%) were hemolysin producer. MRHA is present on twenty-nine strains (35.37%) of eighty-two strains. MRHA positive strains carry a plasmid of 60MDa, a putative factor involved in adherence. This plasmid might be specific for MRHA positive strains. MRHA positive strains were observed in serotype 01, 018, 055, 086a, 0119, 0126, and 0142. Twenty-six strains of E. coli showed three patterns of adherence to HEp-2 cells namely, localized, diffuse, and aggregative adhesion. Twenty-two strains hybridized with the bfpA probe, while all eighty-two strains did not hybridize with the probes, LT, STh, STp. The restriction fragment patterns of chromosomal DNA digested with AotI analysed by PFGE of hemolysin-producing E. coli ten strains were compared with eight different types. Three of E. coli serotype 01, 08 and 0126 showed the same chromosomal DNA fragment patterns.
Blood Cells ; Busan ; DNA ; DNA Probes ; Escherichia coli* ; Escherichia* ; Humans ; Immune Sera ; Plasmids ; Serotyping*

The hydrophobicity assay and DNA probe hybridization assay were compared for analysis of enterotoxigenic Escherichia coli(ETEC), heat-labile enterotoxin(LT) and heat-stable enterotoxin (ST). The ETEC isolated from diarrheal patients were serotyped and investigated for the presence of colonization factor antigens CFA/1, CFA/II, CFA/III and CFA/IV with the expression of mannose-resistant hemagglutination(MRHA) and the levels of surface hydrophobicity. The following results were obtained. 1. Out of these 48 strains, 34 strains were found to be positive for LT production by DNA probe hybridization assay. Out of 34 strains, 1 strain was ST producer, 25 strains were LT producers, and 8 strains were produced both ST+LT producers by DNA probe hybridization assay. 2. Out of 34 strains of positive DNA probe hybridization test, 31 strains was positive in the hydrophobicity test. Among strains of positive hydrophobicity test, 20, 1, and 7 strains produced only LT, only ST and both ST-LT, respectively. Screening efficiency for identifying ETEC by salting out test was 82.4% in sensitivity and 78.6% in specificity. For ETEC detection, the hydrophobicity assay was the least sensitive but was simple, rapid and a good substitute for the DNA probe hybridization assay. 4. CFAs were identified in 43.8% of ETEC strains; 2.1% of the CFAs strains with CFAs harbored CFA/I, 29.2% carried CFA/II, 16.7% carried CFA/III and CFA/IV. And 35.4% expressed none of these CFAs. CFA/I was found in ETEC of serotype 0128: K67, CFA/II was 0128: K67, 0142: K+ and 0159: K+, CFA/III was 086a: K15 and 0128: K67, CFA/IV was 0 86a: K15, 0128: K67, 0125: K70 and 0148: K+.
Colon ; DNA* ; Enterotoxigenic Escherichia coli* ; Enterotoxins ; Escherichia ; Humans ; Hydrophobic and Hydrophilic Interactions* ; Mass Screening ; Sensitivity and Specificity

No Abstract Available.
Methicillin Resistance* ; Methicillin* ; Methicillin-Resistant Staphylococcus aureus* ; Staphylococcus aureus* ; Staphylococcus*

No Abstract Available.
Vibrio parahaemolyticus* ; Vibrio* ; Virulence Factors* ; Virulence*