• Title/Summary/Keyword: Salmonella assay system

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Comparison of Isolation Agar Method, Real-Time PCR and Loop-Mediated Isothermal Amplification-Bioluminescence for the Detection of Salmonella Typhimurium in Mousse Cake and Tiramisu (Mousse cake와 Tiramisu에 인위접종된 Salmonella Typhimurium의 식품공전 분리배지, Real-time PCR과 Loop-mediated isothermal amplification-bioluminescence의 검출 특성 비교)

  • Lee, So-Young;Gwak, Seung-Hae;Kim, Jin-Hee;Oh, Se-Wook
    • Journal of Food Hygiene and Safety
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    • v.34 no.3
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    • pp.290-295
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    • 2019
  • Salmonella spp. are frequently associated with food and are among the most important foodborne pathogens. The recent Salmonella out breaks in Korea was associated with chocolate mousse cakes served with school meals during September 2018. The objective of this research was to compare the 3M Molecular Detection Assay 2 - Salmonella and the Korean Standard Method of Salmonella in artificially inoculated mousse (chocolate and cheese) and tiramisu cakes. Mousse (chocolate and cheese) and tiramisu cakes were artificially inoculated with S. Typhimurium. Twenty five gram of sample was enriched with 225 mL buffered peptone water for incubation at $37^{\circ}C$ for 24 h. After enrichment, the cultures were analyzed by using the 3M Molecular Detection Assay 2 - Salmonella and the Korean Standard Method. Most of the inoculated samples showed similar results except the chocolate mousse cakes, in which real-time PCR was unable to detect S. Typhimurium even after $10^4CFU/25g$ of inoculation. However, S. Typhimurium inoculated at a concentration of $10^0CFU/25g$ was detected by using 3M Molecular Detection Assay 2 - Salmonella. In chocolate mousse, detection of S. Typhimurium using real-time PCR was partially successful when dark chocolate was added at less than 15%. Negative results in real-time PCR and 3M Molecular Detection Assay 2 - Salmonella were confirmed by gel electrophoresis. The data indicated that dark chocolate could inhibit amplification of the target gene in the PCR reactions. In conclusion, the 3M Molecular Detection Assay 2 - Salmonella was better than the Korean Standard Method (real-time PCR) for the detection of S. Typhimurium in chocolate mousse cakes and chocolate mousse.

Mutagenicity of N-Nitrosodimethylamine in Salmonella / Microsome Assay and the Effect of Vitamin C on the Formation of N-Nitrosodimethylamine (Salmonella / Microsome Assay 에서의 N-nitrosodimethylamine의 돌연변이 유발성과 N-nitrosodimethylamine의 생성에 대한 비타민 C의 영향)

  • 김소희;박건영;서명자
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.20 no.3
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    • pp.260-265
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    • 1991
  • The effective method to detect the mutagenicity of N-nitrosodimethylamineI (NDMA) by using Salmonella/microsome assay was studied. The Effect of vitamin C on the mutagenicity of the formed NDMA and during the formation of NDMA from nitrite and secondary amine was also investigated. Aroclor 1254-induced hamster S9 mix was more effective in activation NDMA than rat S9 mix induced by Aroclor 1254 or phenobarbital. Dimethyl sulfoxide and ethanol suppressed the mutagenic effect of NDMA, however, phosphate butter (pH 7.4), distilled water, 95% methanol and Tween 80 + water (1 : 4) were the appropriate dissolving system in the mutagenicity test of NDMA. Vitamin C did not show any inhibitory effect on the mutagenicity of the formed NDMA. However, the revertants of Salmonella typhinutrium TA 100 were significntly reduced (p<0.05) when vitamin C was added to the reaction mixture of nitrite and dimethylamine during the formation of NDMA. The amount of the formed NDMA was analyzed using HPLC and the level was decreased by about 95%. Thus it was concluded that vitamin C inhibited greatly the formation of NDMA from nitrite and dimethylamine.

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Comparison of Gold Biosensor Combined with Light Microscope Imaging System with ELISA for Detecting Salmonella in Chicken after Exposure to Simulated Chilling Condition

  • Mi-Kyung Park
    • Journal of Microbiology and Biotechnology
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    • v.33 no.2
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    • pp.228-234
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    • 2023
  • In this study, the performance of a gold biosensor combined with light microscope imaging system (GB-LMIS) was comparatively evaluated against enzyme-linked immunosorbent assay (ELISA) for detecting Salmonella under simulated chilling condition. The optimum concentration of antiSalmonella polyclonal antibodies (pAbs) was determined to be 12.5 and 100 ㎍/ml for ELISA and GBLMIS, respectively. GB-LMIS exhibited a sufficient and competitive specificity toward three tested Salmonella among only. To mimic a real-world situation, chicken was inoculated with Salmonella cocktail and stored under chilling condition for 48 h. The overall growth of Salmonella under chilling condition was significantly lower than that under non-exposure to the chilling condition (p < 0.05). No significant differences in bacterial growth were observed between brain heart infusion and brilliant green broth during the enrichment period (p > 0.05). Finally, both GB-LMIS and ELISA were employed to detect Salmonella at every 2-h interval. GB-LMIS detected Salmonella with a competitive specificity by the direct observation of bacteria on the sensor using a charge-coupled device camera within a detection time of ~2.5 h. GB-LMIS is a feasible, novel, and rapid method for detecting Salmonella in poultry facilities.

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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Development of Geno-chromatographic Assay System for Salmonella species

  • Kim, Seok-Ha;O, Hong-Il;Jo, Jeong-Hwan;Baek, Se-Hwan
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.827-828
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    • 2001
  • We developed a geno-chromatographic assay system based on hybridization between target DNA (e.g., amplified product from PCR) and a capture probe immobillized on a predetermined site of membrane. This system can offer a rapid detection of a gene sequence specific to a certain type of cells as well as simplicity of the analytical procedure. Such performances were demonstrated by utilizing Salmonella typhimurium as model analyte.

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Rapid Detection for Salmonella spp. by Ultrafast Real-time PCR Assay (Ultrafast Real-time PCR법을 이용한 살모넬라의 신속 검출)

  • Kim, Seok Hwan;Lee, Yu-Si;Joo, In-Sun;Kwak, Hyo Sun;Chung, Gyung Tae;Kim, Soon Han
    • Journal of Food Hygiene and Safety
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    • v.33 no.1
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    • pp.50-57
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    • 2018
  • Salmonella continue to be a major cause of food poisoning worldwide. The rapid detection method of food-borne Salmonella is an important food safety tool. A real-time polymerase chain reaction (PCR) has been used as a rapid method for the detection of pathogens. It has been recently reported that NBS LabChip real-time PCR is a novel, ultrafast, and chip-type-convenient real-time PCR system. We developed the assay method based on NBS LabChip real-time PCR for the rapid detection of Salmonella, which its reaction time was within 20 minutes. Two target genes (invA and stn) were selected to design target specific primers and probes. The new method was validated by checking specificity and sensitivity (limit of detection). This study included forty-two target and twenty-one non-target strains to assess the specificity. This assay was able to identify the 42 Salmonella strains correctly. The limit of detection (LOD) was $10^1copies/{\mu}L$ in Salmonella genomes DNA, while LOD incubated for 4 hr in the inoculated sausage sample ranged from $10^1CFU/g$ to $10^2CFU/g$ as an inoculated cell count. The assay developed in this study could be applied for the investigation of food poisoning pathogens.

Development of One-Step Immuno-Chromatography Assay System for Salmonella typhimurium (Immuno-Chromatography 방법을 이용한 식중독균(Salmonella typhimurium) 1단계 분석시스템의 개발)

  • 백세환;이창우이창섭육순학
    • KSBB Journal
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    • v.11 no.4
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    • pp.420-430
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    • 1996
  • One-step immuno-chromatography assay system for heat-killed Salmonella typhimurium antigens was developed. Three major components used were a glass fiber membrane (placed at the bottom of the system) with an antibody (specific to the analyse, detection antibody)-gold conjugate deposited in a dry state on the surface, a nitrocellulose membrane (middle) with an antibody (also, specific to the analyse but recognized different epitome: capture antibody) and anti-detection antibody immobilized in spatially separated areas, and a cellulose membrane (top) as absorption pad. These membranes were partially superimposed such that a wicking of aqueous solution containing sample can continuously take place through membranes. Variables that affected the system performance were the concentration of capture antibody, the location on the membrane, inert protein used for blocking of the membrane and for carrying the sample, and the concentration of the gold conjugate. Under optimal conditions, within 15 minutes after absorption of a sample solution from the bottom of the system antigen-antibody complexes of sandwich type were formed on the membrane surface area with immobilized capture antibody and a color signal was generated in proportion to the analyse concentration. The minimum do tection limit of the analyse was $1{\times}106$ Salmonella cells/mL.

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Antimutagenic Effect of Plant Flavonoids in the Salmonella Assay System

  • Choi, Jae-Sue;Park, Kun-Young;Moon, Suk-Hee;Rhee, Sook-Hee;Young, Han-Suk
    • Archives of Pharmacal Research
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    • v.17 no.2
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    • pp.71-75
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    • 1994
  • The antimutagenic effects of 27 kinds of plant flavonoids on the mutagenicity of aflatoxin $B_1(AFB_1)$ and N-methyl-N'-nitro-N-nitrosoguanidine(MANG) in Salmonella typhimurium TA 100 were investigated. In the mixed applications of $AFB_1\;(1\;\mu{g/plate)}$ with the flavonoids $(300\;\mu{g/plate)}$ in the presence of a mammalian metabolic activation system (S9 mix), chrysin, apigenin, luteolin and its glucoside, kaempferol, fisetin, morin, naringenin, hesperetin, persicogenin, (+)-catechin and (-)epicatechin showed the antimutagenic effect against $AFB_1$ with more than 70% inhibition rate. A little or no antimutagenicities except flavone against MNNG $(0.5\;\mu{g/plate)}$ were observed. For the antimutagencity of the flavonoids on $AFB_1$, the flavonoid structure that contains the free 5, 7-hydroxyl gorup seemed to be essential. However, saturation of the 2, 3-double bond of elimination of the 4-keto group did not affect the activity.

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Biofilm Formation of Food-borne Pathogens under Stresses of Food Preservation (식품 보존 스트레스에서의 식중독세균의 생체막 생성)

  • Lee, No-A;Noh, Bong-Soo;Park, Jong-Hyun
    • Korean Journal of Food Science and Technology
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    • v.38 no.1
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    • pp.135-139
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    • 2006
  • Most bacteria form biofilm as self-defence system, making efficient food sanitization, preservation, and instrument washing more difficult. Biofilm formation of Salmonella, E. coli, B. cereus, and S. aureus was observed during 24 hr food preservations by performing microtiter plate and glass wool assays. Most cells formed biofilm and attached onto glass wool. When biofilm formation and injury were analyzed on the microtiter plate, 10 and 20% acid-injured E. coli and S. aureus, respectively, 30-50% cold temperature $(4^{\circ}C)-injured$ B. cereus and E. coli, and 30-55% 6% sodium chloride solution-injured Salmonella showed significant biofilm formation. Results indicate biofilm formation level differed within species depending on type of stress.

Antimutagenic and Anticance Effects of Buchu Kimchi

  • Jung, Keun-Ok;Lee, Kyeoung-Im;Suh, Myoung-Ja;Park, Kun-Young
    • Preventive Nutrition and Food Science
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    • v.4 no.1
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    • pp.33-37
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    • 1999
  • The antimutagenic effects of buchu kimchi and Chinese cabbage kimchi and theri cytotoxic effects against human cancer cell line were investigated in the Salmonella typhimurium system and MTT assay, respectively. Leek and Chinese cabbage were aslo evaluated in the same system. Buchu kimchi was fermented at 15 $^{\circ}C$ for 4 days . Buchu kimchi samples showed somewhat higher antimutagenic effects against aflatoxin B1(AFB1) than CHinese cabbage kimchi in Salmonella typhimurium TA100 strain. There was no difference onthe antimutagenic activity according to the length of fermentation . Leek exerted stronger antimutagenicity against AFB1 than Chinese cabbage in the Ames assay. In MTT assay, 6-day fermented buchu kimchin revealed the highest cytotoxicity against AGS human gastric adenocarcinoma cells in which 62% and 82% of the inhibition were observed wiht the addition of 100ug, 400ug/well, respectively. Buchu kimchi samples caused 60~70% inhibition on the proliferation of HT-29 at 400ug/well. Leek exhibited higher antiproliferative effect against both AGS cells and HT-29 cells than Chinese cabbage in MTT assay. From these results, it is considered that buchu kimchi has stronger antimutagenic and in vitro anticancer effects than Chinese cabbage kimchi and the high inhibition rate of buchu kimchi probably results from leek, the major ingredient of buchu kimchi .

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