• Journal of the East Asian Society of Dietary Life
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• v.11 no.1
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• pp.26-32
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• 2001

독소원성 대장균(enterotoxigenic Escherichia coli, ETEC, EC81, serotype O148:H28)이 생산하는 heat labile enterotoxin(LT)를 검색해 본 결과, reversed passive latex agglutination(RPLA)법에 있어서는 2배로 희석한 용액 (50 ng)으로부터 64로 희석한 용액 (1.56 ng)에서까지 양성반응을 보였으며 polymerase chain reaction (PCR)기법에 있어서는 10 ng으로부터 1 pg희석용액에서까지 147-base pair(bp)의 LT DNA fragment가 확인되었다. Clostridium perfringens A형 (NCTC8238, Hobbs serotype 2)이 생산한는 장독소를 검색해 본 결과, RPLA법에 있어서는 2배로 희석한 용액 (50 ng)으로부터 64로 희석한 용액 (1.56 ng)에서 까지 양성반응을 보여 독소원성대장균이 생산하는 LT와 일치하였으나, PCR기법에 있어서는 10ng로부터 10 pg 희석용액에서 까지 354-bp의 DNA fragment가 확인되어 독소원성대장균이 생산하는 LT보다 1/10의 낮은 감도를 보였다. PCR기법은 RPLA법에 비하여 훨씬 신속하고 소량의 sample로 장독소를 확인할 수 있었다.

• Journal of Environmental Health Sciences
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• v.23 no.4
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• pp.45-49
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• 1997

Clostridium perfringens A형이 생산하는 장독소를 검색해본 결과, RPLA법에 있어서는 2배로 희석한 용액으로부터 64배로 희석한 용액 (NCTC 8239 Hobbs serotype 3 CPE$^+$)에서까지 양성반응을 보였으며 PCR 기법에 있어서는 10 pg 희석 용액까지 364 bp의 장독소 DNA fragment(NCTC 8238 Hobbs serotype 2 CPE$^+$)를 확인 할수 있었다. 그러므로 장독소를 검색하기 위해서는 PCR기법이 RPLA법에 비하여 훨씬 감도가 높음을 확인할 수 있었다.

• Korean Journal of Veterinary Research
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• v.39 no.4
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• pp.806-810
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• 1999

A multiplex-polymerase chain reaction (PCR) assay was used to detect staphylococcal enterotoxin production by 12 strains of Staphylococcus aureus isolated from clinical specimens. To evaluate the efficacy and/or sensitivity of this method, the results were compared to those obtained with the reversed passive latex agglutination kit (SET-RPLA, Denka Seiken, Japan). Of 10 strains positive by PCR were positive by RPLA but two strains, representing high sensitivity of the former method. Enterotoxin B was the most prevalent by the two methods. The kappa index between the two methods was 0.826, indicating a higher agreement and fully reliable for use. These results would suggest that sensitive, inexpensive, and relatively rapid multiplex-PCR technique may be an effective means for the detection of staphylococcal enterotoxin genes as an alternative to traditional methods such as kits or immunological methods, which depend upon the amount of enterotoxin produced.

• Korean Journal of Food Science and Technology
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• v.36 no.4
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• pp.683-687
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• 2004

Raw beefs used fer Jangzorim production were evaluated fur contamination of pathogenic bacteria and microorganisms related to spoilage and food safety. Eleven groups of mesophilic, psychrotrophic, and anaerobic microorganisms, and total coliforms were selected to evaluate degree of food contamination. Nine strains including Bacillus cereus, Clostridium botulinum, C. perfringens, and Listeria monocytogenes were selected to evaluate incidences of pathogenic bacteria. Raw beefs harbored large populations of microorganisms, which decreased greatly after heat treatment. Psychrotrophic microorganisms were found to be more abundant than other microorganisms. B. cereus, C. perfringens, and L. monocytogenes were isolated from raw beefs, whereas C. botulinum, Escherichia coli O157:H7, Salmonella spp., Shigella spp., Staphylococcus aureus, and Yersinia enterocolitica were not isolated. Isoiates from Cereus selective agar, clostridium Perfringens agar, and Oxford agar were in 99.8, 99.9 and 98.6% agreements with B. cereus, C. perfringens, and L. monocytogenes at species level, respectively. B. cereus produced enterotoxin with CRET-RPLA method, whereas C. perfringens did not produce enterotoxin with PET-RPLA method.

• Korean Journal of Food Science and Technology
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• v.35 no.1
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• pp.97-102
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• 2003

Raw and washed spinaches were tested to evaluate the incidences of Aeromonas hydrophila, Escherichia coli O157:H7, Plesiomonas shigelloides, Pseudomonas aeruginosa, Salmonella spp., Shigella spp., Yersinia enterocolitica, Bacillus cereus, Campylobacter jejuni, Clostridium perfringens, Listeria monocytogenes, and Staphylococcus aureus. Four pathogenic bacteria were isolated from spinach samples, and identified by morphological and biochemical methods, including API and ATB identification systems. Isolates from MacConkey, Cereus Selective, Clostridium Perfringens, and Baird-Parker agar media were in 99.9, 99.8, 99.9, and 97.8% agreements with A. hydrophila, B. cereus, C. perfringens, and S. aureus at the species level, respectively. SET-RPLA revealed, among the five strains of S. aureus isolates, two produced type A enterotoxin. All five strains of B. cereus isolates produced enterotoxin as revealed with CRET-RPLA.

• Journal of environmental and Sanitary engineering
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• v.20 no.1 s.55
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• pp.23-31
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• 2005

On the purpose of epidemiological survey relate to food poisoning, a total of 114 samples of different salads collected from fast food Restaurants in Gyeonggi-do were for the presence of pathogenic microorganisms. Microbial assessment of salads revealed that TPC($1.1\times10\;\~\;8.4\times10^{5}\;CFU/g$) and coliforms($0\~5.4\times10^{4}\;CFU/g$) exceeded the standards by Solberg et al. ($TPC:10^{5}\;CFU/g,\;coliforms:10^{2}\;CFU/g$). Two pathogenic bacteria were isolated from salad samples, and identified by biochemical methods, including API identification systems. Isolates from PALCAM agar and MYP agar media were in 98.6, $99.8\%$ agreements with Listeria monocytogenes, Bacillus cereus at the species level, respectively. All 7 strains of Bacillus cereus isolates produced enterotoxin as revealed with CRET-RPLA.

• Journal of Microbiology and Biotechnology
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• v.19 no.10
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• pp.1238-1243
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• 2009

The susceptibilities of major enterohemorrhagic Escherichia coli (EHEC) strains to antimicrobial agents and the cytotoxicity of these agents were examined using a total of 38 strains of E. coli O26, O111, and O157, which are the major serogroups of EHEC. Among the 38 strains, 35, 36, and 36 were susceptible to amikacin, imipenem, and norfloxacin, respectively. These antimicrobial agents were further examined to determine their cytotoxicity on Vero cells as well as their effect on the release of Shiga toxins along with trimethoprim/sulfamethoxazole. Each of the E. coli O26, O111, and O157 strains containing both the stx1 and stx2 genes were grown in the absence or presence of these agents at 1/4 minimal inhibitory concentration for 6 h, 12 h, and 18 h. At the concentrations used in this study, none of the agents significantly altered cell count compared with the control group. The level of cytotoxicity in the imipenem group was lower at 12 hand 18 h than their respective controls. In contrast, the level of cytotoxicity in cultures treated with trimethoprim/sulfamethoxazole, norfloxacin, and amikacin was increased. The strains were also examined for the release of Shiga toxins 1 and 2 following treatment with the agents, which were measured by the reversed passive latex agglutination (RPLA) method. The RPLA assay showed a suppression of release of Shiga toxin 2 in the strain cultures containing imipenem. These results indicate that imipenem may be a safe and effective agent for inhibition of these bacteria, which has clinical implications for the treatment of EHEC infections.

• Korean Journal of Clinical Laboratory Science
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• v.37 no.1
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• pp.27-34
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• 2005

Biochemical characteristics, enterotoxin production and antimicrobial susceptibility were determined for 30 strains of Bacillus cereus isolated from stool specimens of diarrhea patients at an university hospital in Chulabuk-do province. Positive rate for VP reaction and citrate utilization were lower, (33 % and 40 % respectively) while the rates of acid production from mannitol, arabinose, and xylose were higher (17 %, 13 % and 3 % respectively) than those obtained by other investigators. The enterotoxin gene was detected in 18 of 30 isolates (60 %) by PCR, and the toxin was detected from all of the toxin gene-positive isolates by RPLA test. The agar dilution test showed that all isolates were resistant to penicillin G and 73 % were to cephalothin, but all were susceptible to ciprofloxacin, clindamycin, erythromycin, fusidic acid, gentamicin, rifampin, teracycline and vancomycin. We conclude that B. cereus isolates producing acid from mannitol, arabinose and xylose exist, that PCR can be used to detect enterotoxin genes rapidly and accurately, and that this organism is susceptible to various antimicrobial agents though not penicillin G and cephalothin.

• Journal of Microbiology and Biotechnology
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• v.25 no.9
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• pp.1460-1466
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• 2015

In this study, the prevalence of Shiga toxin-producing Escherichia coli (STEC) was investigated among raw meat or meat products from slaughterhouses and retail markets in South Korea, and their potential for antibiotic resistance and virulence was further analyzed. A total of 912 raw meats, including beef, pork, and chicken, were collected from 2008 to 2009. E. coli strains were frequently isolated in chicken meats (176/233, 75.9%), beef (102/217, 42.3%), and pork (109/235, 39.2%). Putative STEC isolates were further categorized, based on the presence or absence of the Shiga toxin (stx) genes, followed by standard O-serotyping. Polymerase chain reaction assays were used to detect the previously defined virulence genes in STEC, including Shiga toxins 1 and Shiga toxin 2 (stx1 and 2), enterohemolysin (ehxA), intimin (eaeA), STEC autoagglutination adhesion (saa), and subtilase cytotoxin (subAB). All carried both stx1 and eae genes, but none of them had the stx2, saa, or subAB genes. Six (50.0%) STEC isolates possessed the ehxA gene, which is known to be encoded by the 60-megadalton virulence plasmid. Our antibiogram profiling demonstrated that some STEC strains, particularly pork and chicken isolates, displayed a multiple drug-resistance phenotype. RPLA analysis revealed that all the stx1-positive STEC isolates produced Stx1 only at the undetectable level. Altogether, these results imply that the locus of enterocyte and effacement (LEE)-positive strains STEC are predominant among raw meats or meat products from slaughterhouses or retail markets in Korea.

• Korean Journal of Food Science and Technology
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• v.36 no.1
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• pp.44-49
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• 2004

Changes in quality properties of Kochujang prepared with Paecilomyces japonica powder and extract using different solvents were investigated during 90 days of fermentation at $20^{\circ}C$. Although moisture contents were not significantly different, pH of P. japonica-added Kochujang was lower than that of control group without P. japonica, and decreased with increasing fermentation time. Amino nitrogen content increased up to 60 days of fermentation and decreased slightly after 90 days, with that of P. japonica-added Kochujang showing highest on 30 and 60 days at 179.2 and 282.2 mg%, respectively, higher than control gruup. L, a, and b values decreased in proportion to fermentation period, with P. japonica-added Kochujang, particularly P. japonica powder-added Kochujang, lower than those of control g개up. Sensory evaluation test showed color of control group was 'clear red', whereas that of P. japonica powder-added Kochujang was 'dark reddish brown' and P. japonica extract-added Kochujang was darker than control group; consumer preference for dark color was low, Textures of all samples were 'glossy and smooth', showing high consumer preference. Salt content of P. japonica-added Kochujang was higher than that of control group, with P. japonica extract-added Kochujang higher than that made with powder Hot taste or P. japonica-added Kochujang was weaker, whereas its flavor higher, than control group, with P. japonica powder-added Kochujang showing highest flavor score. Overall preference was higher for P. japonica-added Kochujang than control group, with P. japonica water extract-added Kochujang showing the highest score.