• Title/Summary/Keyword: microplate method

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Prevalence and Characterization of Enterohemorrhagic Escherichia coli (EHEC) Isolated from Ground Beefs Distributed in Gyeong-In Region (경인지역에 유통되는 분쇄육 중 장출혈성대장균의 분포 조사 및 특성 연구)

  • Kim, Eun-Jeong;Park, Yong-Chjun;Cho, Joon-Il;Lee, Jong-Ok;Kim, Hee-Yun
    • Korean Journal of Food Science and Technology
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    • v.38 no.6
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    • pp.773-778
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    • 2006
  • The objective of this study was to evaluate three verocytotoxin-producing Escherichia coli (VTEC) detection kits to detect the presence of VT genes: Doupath Verocytotoxin (GLISA) developed by MERCK, ProsPect Shiga Toxin E. coil (STEC) Microplate Assay (ELISA) developed by Remel, and a polymerase chain reaction method. Our laboratory verified artificially inoculated samples. All three methods could detect very low numbers of VTEC, but VT-PCR had the best sensitivity for VTEC detection. From April through September 2005, 257 ground-beefs from supermakets and traditional markets were examined for the presence of VTEC by polymerase chain reaction immediately after purchase and total viable counts (TVC) were determined. VTEC was isolated from 30 of 257 ground-beefs. A variety of serogroups was found, including 10 stains belonging to the virulence type EHEC, but major serogroups such as O157, O26 and O111 were nor found.

Estimating dehalogenation reactivity of nanoscale zero-valent iron by simple colorimetric assay by way of 4-chlorophenol reduction

  • Mines, Paul D.;Kaarsholm, Kamilla M.S.;Droumpali, Ariadni;Andersen, Henrik R.;Hwang, Yuhoon
    • Environmental Engineering Research
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    • v.25 no.2
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    • pp.197-204
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    • 2020
  • A number of different nanoscale zero-valent iron (nZVI) materials have been prepared and compared depending on the desired properties for the particular application, but different physicochemical properties of this prepared nZVI make it difficult to universally compare and standardize them to the same scale. In this study, we aimed to demonstrate a simple microplate-based colorimetric assay using 4-chlorophenol as an indicator with respect to the remediation of real treatment targets, such as trichloroethylene (TCE), 1,1,1-trichloroethane (TCA), and atrazine. Effect of nickel contents on 4-chlorophenol reduction was successfully investigated by the miniaturized colorimetric assay. In the same manner, the effect of nickel contents on dehalogenation of TCE, TCA, and atrazine was investigated and the pseudo-first-order kinetic constants were compared with the results for 4-chlorophenol. The similar pattern could be observed between 4-chlorophenol reduction obtained by colorimetric assay and TCE, TCA, atrazine reduction obtained by a traditional chromatographic method. The reaction kinetics does not match perfectly, but the degree of reaction can be estimated. Therefore, the colorimetric assay can be a useful and simple screening tool to determine nZVI reactivity toward halogenated organics before it is applied to a particular remediation site.

Studies on serological tests for pullorum disease (추백리의 혈청학적 진단법에 관한 연구)

  • 김정태;심항섭;김태종;고태오;우종태;유기승;박유순
    • Korean Journal of Veterinary Service
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    • v.21 no.3
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    • pp.313-323
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    • 1998
  • In order to establish a sensitive and specific diagnostic method for detection of antibody to Salmonella pullorum, a enzyme-linked immunosorbent assay(ELISA) was designed and standardized. The diagnostic efficacy of the established ELISA was compared with that of the serum plate agglutination test and immunodiffusion test for pullorum disease. 1. The chicken hyperimmune sera to Salmonella pullorum, S gallinarum, S typhimurium and S typhi were shown the cross reaction to S pullorum antigen by serum plate agglutination test. 2. When compared the cross reaction titer of microplate agglutination test for chickens hyperimmune sera, it was found that the titer were 64 in S pullorum, 32 in S gallinarum, 4 in S typhimurium and 8 in S typhi, respectively. 3. When compared the specificity of various antigen(HA, EA, PA and SA) by the immunodiffusion test, the most suitable antigen was phenol-treated bactrium. 4. The optimal concentration of S pullorum antigen for ELISA was 1 : 160 dilution of bacterium. 5. The efficacy of the ELISA for detection of S pullorum antibody was compared with serum Plate agglutination test and immunodiffusion test in chickens infected with S pullorum. The antibody was first detected at 6 days after infection using three tests examined. The antibody was alldetected at 9 days by ELISA, at 12 days by serumplate agglutination test, at 15 days by immunodiffusion test.

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Prevalence of toxoplasma antibody in swine by microplate latex agglutination test in Kyeongnam district (Latex 응집반응에 의한 경남중부지역 돼지의 톡소플라즈마병 항체조사)

  • Lee, Byung-hoon;Kim, Cha-yong;Suh, Myung-deuk
    • Korean Journal of Veterinary Research
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    • v.33 no.2
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    • pp.263-268
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    • 1993
  • This study was conducted to detect the serum antibody of toxoplasma in swine from breeding-pig, rearing-pig farm and slaughtered pig in abattior by latex agglutination(LA) test. The perfomance of LA test was carried out with commercial Toxo-MT kit(Eiken Chemical Co.)by Tsubota and Ozawa's method. The cut-off titer of positive and negative reactions by Toxo-MT antigen used in this experiment was determined as the serum titer of 1 : 32. Positive rate of toxoplasma antibody from the total of 823 serum samples by LA test was 17.0%(140 cases). And positive rates of toxoplasma antibody against serum samples of 194 from breeding-pig farm, 273 from rearing-pig farm and 356 from abattior were 91 cases(46. 9%), 23 cases(8.4%) and 26 cases(7.3%), respectively. The distributions of serum antibody titers in 823 test sera by LA test were shown 51 cases(36.3%) in 1:32, 40(28.6%) in 1:64, 17(12.1%) in 1:128, 14(10.0%) in 1:256, 3(2.1%) in 1:512, 5(3.6%) in 1:1024 and 3(2.1%) in 1:2048. The ranges of positive rate from the sera in each group of breeding-pig farms were 20~61.9%.

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Optimization of Tri-enzyme Extraction Procedures for the Microbiological Assay of Folate in Red Kidney Bean and Roasted Peanut Using Response Surface Methodology

  • Choi, Young-Min;Eitenmiller, Ronald R.;Kim, Seon-Hee;Lee, Jun-Soo
    • Food Science and Biotechnology
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    • v.18 no.1
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    • pp.31-35
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    • 2009
  • Total folate content was determined by microbiological assay using Lactobacillus casei spp. rhamnosis (ATCC 7469) with a 96-well microplate technique. Using roasted peanut and red kidney beans as representative legume samples, response surface methodology (RSM) was supplied to optimize the trienzyme procedures for the determination of folate in legumes. After response surface regression (RSREG), the second-order polynomial equation was fitted to the experimental data. Ridge analysis showed that the optimal digestion times were <2 hr for $Pronase^{(R)}$ and $\alpha$-amylase, and <5 hr for conjugase to obtain maximal folate values for legume samples. This study confirms that established digestion times for cereal products (AOAC Method 2004.05) of 3 for protease and 2 hr for $\alpha$-amylase are applicable to legumes. Conjugase treatment can be reduced to 5 from 16 hr and the conjugase level to 5 from 20 mg per sample, providing significant cost saving.

Synthesis, Docking Study and In-vitro Evaluation of Anti-Tuberculosis Activity of Tri Substituted Imidazoles Containing Quinoline Moiety

  • Sahana, S.;Vijayakumar, G.R.;Sivakumar, R.;Sriram, D.;Saiprasad, D.V.
    • Journal of the Korean Chemical Society
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    • v.66 no.3
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    • pp.194-201
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    • 2022
  • A simple, efficient, and cost-effective method has been employed for the synthesis of 2,4,5-trisubstituted imidazole derivatives (3a-j) containing quinoline substituent at 2nd position. Title compounds were obtained by multicomponent reaction (MCR), involving aryl substituted 1,2-diketone, quinoline carbaldehyde and ammonium acetate in the presence of acetic acid solvent under mild reaction conditions. The newly synthesized quinoline containing imidazole derivatives were confirmed through FT-IR, 1H-NMR, 13C-NMR and mass spectral analysis. In-vitro microplate alamar blue assay (MABA) to determine the MIC (minimum inhibitory concentration) values against Mycobacterium tuberculosis H37Rv was performed for the synthesized compounds. The synthesized compounds exhibited activity against Mycobacterium tuberculosis and among which compounds, 3d, 3f and 3i showed good activity. The highest activity was showed with compound 3i. The anti-mycobacterial activity results are well correlated with the computational molecular docking analysis, which was performed for the synthesized compounds prior to the evaluation of the activity.

Acetylcholinesterase Inhibitory and Antioxidant Properties of Aster yomena Extract (쑥부쟁이 추출물의 아세틸콜린에스테라제 저해 및 항산화 활성)

  • Bae, Jong-Sup;Kim, Tae-Hoon
    • The Korea Journal of Herbology
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    • v.24 no.4
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    • pp.121-126
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    • 2009
  • Objectives : To evaluate the radical scavenging and acetylcholinesterase (AChE) inhibitory activities of the ethylacetate (EtOAc)-soluble portion of a methanolic extract of Aster yomena, three different assay systems were performed. Methods : The antioxidant activity of A. yomena extract was tested as its capacity to scavenging free radicals of DPPH and $ABTS^+$, which has been widely used to evaluate the antioxidant activity of natural products from plant sources. AChE inhibitory activity was tested against mouse brain AChE by spectrophotometric method of Ellman using ELISA microplate reader. Results : The methanolic extract of A. yomena was fractionated and the EtOAc-soluble portion showed significant AChE inhibitory and free radical scavenging effects. Also the EtOAc-soluble portion revealed the highest phenolic contents as compared to the other extracts. Conclusions : These results indicate that phenolic compounds may be important constituents that give rise to the anti-AChE and antioxidative activities of A. yomena extract. Further phytochemical studies on this plant, for nutraceutical or pharmaceutical application, are warranted.

A Highly Sensitive Indirect Enzyme-Linked Immunosorbent Assay (ELISA) Based on a Monoclonal Antibody Specific to Thermal Stable-Soluble Protein in Pork Fat for the Rapid Detection of Pork Fat Adulterated in Heat-Processed Beef Meatballs

  • Sol-A Kim;Jeong-Eun Lee;Dong-Hyun Kim;Song-min Lee;Hee-Kyeong Yang;Won-Bo Shim
    • Food Science of Animal Resources
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    • v.43 no.6
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    • pp.989-1001
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    • 2023
  • Processed foods containing pork fat tissue to improve flavor and gain economic benefit may cause severe issues for Muslims, Jews, and vegetarians. This study aimed to develop an indirect enzyme-linked immunosorbent assay (iELISA) based on a monoclonal antibody specific to thermal stable-soluble protein in pork fat tissue and apply it to detect pork fat tissue in heat-processed (autoclave, steam, roast, and fry) beef meatballs. To develop a sensitive iELISA, the optimal sample pre-cooking time, coating conditions, primary and secondary dilution time, and various buffer systems were tested. The change in the iELISA sensitivity with different 96-well microtiter microplates was confirmed. The detection limit of iELISA performed with an appropriate microplate was 0.015% (w/w) pork fat in raw and heat-treated beef. No cross-reactions to other meats or fats were shown. These results mean that the iELISA can be used as an analytical method to detect trace amounts of pork fat mixed in beef.

Epidemiological characteristics on fowl typhoid outbreak in Kyongnam province and comparison of diagnostic methods for identification of salmonella gallinarum (경남지역에서 발생한 가금티푸스의 역학적 특성 및 진단방법에 대한 비교 시험)

  • 최유정;김도경;김용환
    • Korean Journal of Veterinary Service
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    • v.23 no.4
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    • pp.349-360
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    • 2000
  • An epidemiological survey was conducted to investigate fowl typhoid outbreaks in Kyungnam province of Korea. The causative agent, salmonella gallinarum was isolated from 68 chicken samples of tentatively diagnosed fowl typhoid cases occurred during the period from January 1996 to September 1999. Comparative studies were also carried out to evaluate the diagnostic methods for detection of S gallinam The results obtained were as follows; 1. Of the 68 cases of tentatively diagnosed fowl typhoid, 56 (82%) cases were determined as fowl typhoid by biochemical test and pathological findings. The other 12 (18%) cases were determined as paratyphoid. 2. Fowl typhoid outbreaks occur continuously all seasons in the year, however the incidence was remarkably increased from May to September. 3. The frequency of incidence of fowl typhoid in terms of regional distribution was relatively high in egg-laying hens facilities, and the mode of transmission is likely to be either egg-to-egg or lateral transfer by wild birds or rats. 4. All of 18 isolates from 56 cases were identified as S gallinarum by biochemical and serological test. 5. Antimicrobial drug susceptibility test against 18 isolates showed that the isolates were highly susceptible to ASH, CZ, CF and GM (above 90%), whereas those strains were 100% resistant to EM, NA and PC. 6. S gallinarum rfbS gene was targeted to be amplified by PCR for comparative detection of S gallinarum in the experimentally infected chickens. The amplified 720bp DNA fragment, which is specific in D serogroup strains of S enterica subspecies was confirmed by agarose gel electrophoresis. 7. A comparison made between fecal culture and PCR-method revealed that later-method was relatively higher in detection rate than that of former method for S gallinarum. 8. Comparison of currently applied methods, rapid serum agglutination test (RST) and microplate agglutination test (MAT), with experimentally infected chickens were made to evaluate sensitivity of detection by neutralizing antibody titration. Both methods detected neutralizing antibodies from the challenged chickens of 5 day post infection. However, positive reactions were determined after 7 and 9 days post infection by MAT and RST, respectively.

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Antimicrobial Effects of Natural Flavonoids and a Novel Flavonoid, 7-O-Butyl Naringenin, on Growth of Meat-borne Staphylococcus aureus Strains

  • Moon, Sun- Hee;Lee, Kyoung-Ah;Park, Keun-Kyu;Kim, Kee-Tae;Park, Yong-Sun;Nah, Seung-Yeal;Mendonca, Aubrey F.;Paik, Hyun-Dong
    • Food Science of Animal Resources
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    • v.31 no.3
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    • pp.413-419
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    • 2011
  • The antimicrobial effects of the natural flavonoids kaempferol, quercetin, apigenin, and naringenin as well as a novel flavonoid 7-O-butyl naringenin against the growth of four meat-born Staphylococcus aureus strains were evaluated. First, the flavonoids were screened for inhibitory effects against the growth of each strain using the paper disc diffusion method. Second, the growth inhibitory effects of flavonoids that showed antimicrobial activity were measured using the microplate method. Third, the bactericidal effects of flavonoids were evaluated in a 0.8% (w/v) NaCl solution. All flavonoids showed bacteriostatic effects at >20 mM. Among the flavonoids studied, quercetin was more effective than the others tested. However, the inhibitory effect of 7-O-butyl naringenin on growth of S. aureus KCCM 32395 was greater than that of quercetin at the same concentration. Additionally, 7-O-butyl naringenin exhibited significant bactericidal effects at >25 ${\mu}M$. When bacterial cells were examined using scanning electron microscopy, it appeared that the S. aureus membranes were damaged or morphologically changed when treated with quercetin and 7-O-butyl naringenin at 200 ${\mu}M$.