• Title/Summary/Keyword: Salmonella assay

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Detection of Escherichia coli O157:H7, Salmonella spp., Staphylococcus aureus and Listeria monocytogenes in Kimchi by Multiplex Polymerase Chain Reaction (mPCR)

  • Park, Yeon-Sun;Lee, Sang-Rok;Kim, Young-Gon
    • Journal of Microbiology
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    • v.44 no.1
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    • pp.92-97
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    • 2006
  • We developed an mPCR assay for the simultaneous detection, in one tube, of Escherichia coli O157:H7, Salmonella spp., Staphylococcus aureus and Listeria monocytogenes using species-specific primers. The mPCR employed the E. coli O157:H7 specific primer Stx2A, Salmonella spp. specific primer Its, S. aureus specific primer Cap8A-B and L. monocytogenes specific primer Hly. Amplification with these primers produced products of 553, 312, 405 and 210 bp, respectively. All PCR products were easily detected by agarose gel electrophoresis, and the sequences of the specific amplicons assessed. Potential pathogenic bacteria, in laboratory-prepared and four commercially available kimchi products, were using this mPCR assay, and the amplicons cloned and sequenced. The results correlated exactly with sequences derived for amplicons obtained during preliminry tests with known organisms. The sensitivity of the assay was determined for the purified pathogen DNAs from four strains. The mPCR detected pathogen DNA at concentrations ranging from approximately 0.45 to $0.05\;pM/{\mu}l$. Thus, this mPCR assay may allow for the rapid, reliable and cost-effective identification of four potentially pathogens present in the mixed bacterial communities of commercially available kimchi.

Antimutagenic Effect and Active Compound Analysis of Kale Juice in Salmonella Assay System (Salmonella 실험계에서 케일즙액의 항돌연변이의 효과와 활성획분)

  • 이선미;박건영;이숙희
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.5
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    • pp.965-971
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    • 1997
  • Antimutagenic effects of kale juice on the mutagenicity induced by $B_{1}(AFB_{1})$ N-methyl-N'-N-nitrosoguanidine(MNNG) in Salmonella assay system were studied. The freeze dried kale juice significantly reduced the mutagenicity induced by $AFB_{1}$ in Salmonella typhimurium TA100 and TA98. However, the kale juice exhibited less inhigbitory effect on the mutagenicity induced by MNNG as the concentrations of the juice sample increased. Also, kale juice after dialysis (>12,000, Mw) appeared to have 42.3∼89.5% of inhibitory effects against $AFB_{1}$, however, the dialyzate did not show any inhibitory effect against MNNG. To separate and identify the antimutagenic compounds from the kale juice, the dialyzates were further fractioned by using Sepharose CL-6B-200 gel filtration. Fraction number 13 showed the strong antimutagenic activity against $AFB_{1}$, and the fraction exhibited positive results of a characterized colour reactions of protein, carbohydrate and phenolic compound. Therefore, one of the possible active compounds from the kale juice was supposed to a glycoprotein(Mw. 270,000) which seemed unstable with heating.

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Genotoxicily Studies of An Anticancer Agent of Camptothecin Series, CKD-602 (Camptothecin계 항암제 CKD-602의 유전독성평가)

  • 하광원;오혜영;허옥순;박장환;손수정;한의식;김종원;강일현;강혁준
    • Environmental Mutagens and Carcinogens
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    • v.18 no.2
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    • pp.129-134
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    • 1998
  • To evaluate the genotoxicity of CKD-602, an anticancer agent the in viかo reverse mutation assay using Salmonella typhimurium, the Chromosome aberration assay using Chinese hamster lung (CHL) cells and the in vivo micronucleus assay using bone marrow cells of ddY mice were performed. In the reverse mutation assay, CKD-602 did not induced mutagenicity in Salmonella typhimurium TA 98, TA 100, TA 1535, and TA 1537 strains with and without metabolic activation. In the chromosome aberration test using CHL cells, there was an increased incidence of structural aberrations induced by CKD-602 without metabolic activation during 24 and 48 hours, but CKD-602 did not induce chromosome aberration with metabolic activation. The in vivo induction of micronuclei was measured in polychromatic erythrocytes of bone marrow of male ddY mice. At 24 hours after treatment with CED-602 by i.p. once, there was an increased incidence of micronucleated polychromatic erythrocytes in bone marrow of ddY male mice.

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Genotoxicify Studies of on Antihypertensive Agent, SKP-450 (고혈압 치료제 SKP-450의 유전독성평가)

  • 하광원;오혜영;박장환;허옥순;손수정;한의식;류근호;조용백
    • Environmental Mutagens and Carcinogens
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    • v.18 no.2
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    • pp.123-128
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    • 1998
  • To evaluate the genotoxicity of SKP-450, an antihypertensive agent the in vitro reverse mutation assay using Salmonella typhimurium, the Chromosome aberration assay using Chinese hamster lung (CHL) cells and the in vivo micronucleus assay using bone marrow cells of ddY mice were performed. In the Reverse mutation test, SKP-450 did not induced mutagenicity in Salmonella typhimurium TA 98, TA 100, TA 1535 and TA 1537 with and without metabolic activation. In the chromosome aberration assay using CHL cells, there was no increased incidence of structural and numerical aberrations with and without metabolic activation. The in vivo induction of micronuclei was measured in polychromatic erythrocytes of bone marrow of male ddY mice at 30 hours after treatment with SKP-450 by p.o once. The results showed no increased incidence of micronucleated polychromatic erythrocytes in bone marrow of ddY male mice treated with SKP-450.

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Detection of Pathogenic Salmonella Using a Surface Plasmon Resonance Biosensor (표면플라즈몬공명 바이오센서를 이용한 살모넬라 검출)

  • Cho, Han-Keun;Kim, Gi-Young;Kim, Woon-Ho;Sung, Min-Sun
    • Journal of Biosystems Engineering
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    • v.35 no.2
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    • pp.116-123
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    • 2010
  • Rapid detection of foodborne pathogens has been a major challenge for the food industry. Salmonella contamination is well known in all foods including pasteurised milk. The possibility of specific detection of Salmonella Enteritidis by surface plasmon resonance (SPR) biosensor was explored using a commercially available portable SPR sensor. Self assembly technique was adopted to immobilize anti-Salmonella antibodies on the gold sensing surface of the SPR sensor. The concentration of polyclonal antibody for use in the SPR biosensor was chosen to 1.0 mg/mL. Experiments were conducted at near real-time with results obtained for one SPR biosensor assay within 1 hour. The limit of detection for Salmonella Enteritidis was determined to be $10^6$ CFU/mL in both PBS buffer and milk samples. The assay sensitivity was not significantly affected by milk matrix. Our results showed that it would be possible for employing the SPR biosensor to detect Salmonella Enteritidis in near real-time.

Evaluation of Mutagenicity with Gamgung-tang Using Host-Mediated Assay (Host-Mediated Assay를 이용한 감궁탕의 돌연변이원성 평가)

  • Shon, Yun-Hee;Kim, Cheorl-Ho;Nam, Kyung-Soo
    • Korean Journal of Pharmacognosy
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    • v.36 no.2 s.141
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    • pp.93-96
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    • 2005
  • Mutagenicity of Gamgung-tang (GGT) was tested using in vitro S-9 mixture in vitro host-mediated assay with Salmonella typhimurium. In the previous reports, GGT was tested for the safety using Ames(-S-9), Bacillus subtilis Rec, and umu gene expression mutagenicity tests. Mutagenic activity in any assays we tested was not found. In this report, we further investigated safety of GGT after metabolic activation in vivo. Ames test with S-9 mixture and host-mediated assay with Salmonella typhimurium TA98 were used to identify metagenic property of GGT. GGT was administered 3 times with i.m. to Balb/c mice did not induced mutagenic effect in Salmonella typhimurium TA98 recovered from the liver after 3.5h with i.p. treatment. Over the entire dose range $(3{\sim}150mg/mouse)$ tested no toxicity was detected to the bacterial cells. These results suggest that there was no DNA damage and mutagenicity by GGT.

Mutagenecity Test of SDK (SDK시제품(가칭)에 대한 변이원성시험)

  • 정지윤;이원우;임종희;남정석;제정환;이광훈;강병철;이병희;박재학
    • Toxicological Research
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    • v.14 no.2
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    • pp.211-216
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    • 1998
  • In order to evaluate the mutagenic potential of SDK(skin decontamination kit) produced by Agency for Defense Development(ADD), were performed Salmonella typhimurium reversion assay, chromosomal aberration test on chinese hamster ovarian cells and in vivo micronucleus assay using mouse bone marrow cells according to the established regulation of Korean Food and Drug Administration. In the reverse mutation test using Salmonella typhimurium TA98, TA100, TA1535 and TA1537 did not in-crease the number of revertant at any of the concentration tested in this study. SDK did not increase the number of cells having structural or numerical chromosome aberration in cytogenetic test. In mouse micronucleus test, no significant increase in the occurrence oj micro nucleated polychromatic erythrocytes were observed in ICR male mice intraperitoneally administered with SDK. These results indicate that SDK has no mutagenic effects under these experimental conditions.

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A Comparison of Inhibitory Effects in Brown and White Rice ( Oryza sativa L. ) against Mutagenicity Induced by Tryptophan Pyrolysates (트립토판 열분해물 유발 변이원성에 대한 현미 및 백미의 억제 효과 비교)

  • Jo, Jeong-Sun;Yu, Jeong-Eun;Jeon, Hyang-Suk
    • Journal of the Korean Dietetic Association
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    • v.3 no.2
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    • pp.105-111
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    • 1997
  • The inhibitory effect of rice(Oryza sartiva L., illpumbyeo) against mutagenicity induced by tryptophan pyrolysates were investigated using Salmonella typhimurium reversion assay. Both methanol extracts of obtained from brown and white rice were found to possess strong activites of inhibiting the mutagenicities of 3-amino-1,4-dimethyl-5H-pyriod[4,3-b]indol(Trp-P-1) and 3-amino-1-methyl-5H-pyrido[4,3-b]indol(Trp-P-2) on Salmonella typhimurium reversion assay. As the concentration of methanol extract increased, inhibitory effect on mutagenicity increased but reached at steady state as inhibition rate of 90% when the concentration was above 10mg/plate. There was no significant difference(p>0.05) in inhibitory effect of methanol extracts between brown and white rice against tryptophan pyrolysates.

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Antioxidative Stress and Antimutagenic Effects of Lentinus edodes Ethanol Extracts (표고버섯 에탄올 추출물의 산화적 스트레스 억제 효과와 항돌연변이 효과)

  • Oh, Se-In;Lee, Mee-Sook
    • The Korean Journal of Food And Nutrition
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    • v.20 no.4
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    • pp.341-348
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    • 2007
  • The antioxidative effect and antimutagenic capacity in ethanol extracts of Lentinus edodes were studied for suggestion of prevention and dietetic treatment of chronic diseases and development of antioxidative and antimutagenic functional food by employing biological and biochemical assay. The $IC_{50}$ of MDA with BSA conjugation reaction, lipid peroxidation and scavenging effect on DPPH radical in ethanol extracts of Lentinus edodes showed 74.58 mg/assay, 5.747 mg/assay and 0.939 mg/assay respectively. So, the most effective antioxidative capacity in ethanol extracts of Lentinus edodes was the scavenging effect on DPPH radical, among the method used this study. The indirect and direct antimutagenic effects of ethanol extracts of Lentinus edodes were examined by Ames test using Salmonella typimurium TA98 and TA100. The inhibition rates on indirect mutagenicity mediated by 2-anthramine showed 91.67% in the Salmonella typimurium TA98 and 96.60% in the Salmonella typimurium TA100. The inhibitory effect on direct mutagenicity mediated by sodium azide in Salmonella typimurium TA100 was 22.83%. and mediated by 2-nitrofluorene in Salmonella typimurium TA98 was 5.34%. This data indicates that ethanol extracts of Lentinus edodes have more effective effects on indirect mutagenicity than direct mutagenicity. From this result, it believed to have a possible antioxidative and antimutagenic capacities, and taken for the candidate of prevention and dietetic treatment of chronic diseases and development of antioxidative and antimutagenic functional food.

Quantitative Detection of Salmonella typhimurium Contamination in Milk, Using Real-Time PCR

  • JUNG SUNG JE;KIM HYUN-JOONG;KIM HAE-YEONG
    • Journal of Microbiology and Biotechnology
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    • v.15 no.6
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    • pp.1353-1358
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    • 2005
  • A rapid and quantitative real-time PCR was developed to target the invasion A (invA) gene of Salmonella spp. We developed quantitative standard curves based on plasmids containing the invA gene. Based on these curves, we detected Salmonella spp. in artificially contaminated buffered peptone water (BPW) and milk samples. We were able to determine the invA gene copy number per ml of food samples, with the minimum detection limit of $4.1{\times}10^{3}$ copies/ml of BPW and $3.3{\times}10^{3}$ copies/ml of milk. When applied directly to detect and quantify Salmonella spp. in BPW and milk, the present real-time PCR assay was as sensitive as the plate count method; however, copy numbers were one to two logs higher than the colony-forming units obtained by the plate count methods. In the present work, the real-time PCR assay was shown to significantly reduce the total time necessary for the detection of Salmonella spp. in foods and to provide an important model for other foodborne pathogens.