• Title/Summary/Keyword: assay method

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Screening of New Bioactive Materials from Microbial Extracts of Soil Microorganism (I) Antimicrobial Activity from 200 Sampled Using Microdilution Assay

  • Jung, Sang-Oun;Kim, Joon;Chang, Il-Moo;Ryu, Jae-Chun
    • Archives of Pharmacal Research
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    • v.21 no.3
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    • pp.278-285
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    • 1998
  • The microdilution assay recommended by NCCLS (National Committee for Clinical Laboratory Standards) is one of the standardized methods of antibiotic susceptibility test. This method has been widely used clinically to obtain MIC values of antibiotics on pathogenic microorganisms. It is more convenient, rapid and simple to test many samples than other test methods such as agar diffusion assay and broth macrodilution assay. The screening of antimicrobial agents from microbial extracts is too laborious in its process. Therefore, a number of screening methods having more simple procedure have been developed. In our laboratory, we applied microdilution assay for screening the antimicrobial agents. This assay showed dose-response results and was more sensitive than disc diffusion assay in our system. We tested 200 samples of microbial extracts originated from 100 microbial strains and selected several samples as potential candidates. In this report, we show that the microdilution assay is more convenient method in screeing of antibiotic susceptibility than those previously reported.

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Clinical Usefulness of Helicobactor pylori Ag Stool Test (Immunochromatographic Assay) for Diagnosis of H. pylori Infection (Helicobacter pylori 감염진단에 있어 H. pylori Ag Stool 검사 (면역크로마토그라피법)의 임상적 유용성)

  • Seo, Seol
    • Korean Journal of Clinical Laboratory Science
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    • v.42 no.1
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    • pp.38-45
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    • 2010
  • The aim of this study was to assess the Clinical Usefulness of Helicobacter pylori Stool Antigen (HpSA) immunochromatographic assay for the diagnosis of H. pylori infection. In this study, we had compared HpSA-immunochromatographic assay with CLO test and UBT test. From a total of 140 patients (M:F=88:52) with upper endoscopy, biopsy specimens were obtained for CLO test. Stool specimens was collected from all patients and tested using a HpSA-immunochromatic assay. H. pylori infection status was defined as infected if the results of both CLO test and UBT test were positive. CLO test and UBT test findings showed that 92 patients were H. pylori positive and 48 patients were H. pylori negative. According to this definition, the sensitivity, specificity, and positive or negative predictive value (PPV, NPV) of HpSA-immunochromatographic assay were 97.8%, 100%, 100%, and 96%, respectively. Cross reactivity test of HpSA-immunochromatographic assay were performed with 10 enteric bacteria strains in fecal habitat, and there were no false positive reaction. We evaluated the usefulness of HpSA assay for eradication therapy with 10 of 92 H. pylori positive patients, positive results of them at pre-eradication therapy were converted to negative at post-eradication. The HpSA-immunochromatographic assay is a highly sensitive and specific non-invasive diagnostic method for detection of H. pylori infection, a useful diagnostic method for H. pylori in post eradication stage.

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Head-to-Head Comparison between Xpert MTB/RIF Assay and Real-Time Polymerase Chain Reaction Assay Using Bronchial Washing Specimens for Tuberculosis Diagnosis

  • Son, Eunjeong;Jang, Jinook;Kim, Taehwa;Jang, Jin Ho;Chung, Jae Heun;Seol, Hee Yun;Yeo, Hye Ju;Yoon, Seong Hoon;Lee, Seung Eun;Cho, Woo Hyun;Kim, Yun Seong;Jeon, Doosoo
    • Tuberculosis and Respiratory Diseases
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    • v.85 no.1
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    • pp.89-95
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    • 2022
  • Background: With the introduction of Xpert MTB/RIF assay (Xpert), its incorporation into tuberculosis (TB) diagnostic algorithm has become an important issue. The aim of this study was to evaluate the performance of the Xpert assay in comparison with a commercial polymerase chain reaction (PCR) assay. Methods: Medical records of patients having results of both Xpert and AdvanSure TB/NTM real-time PCR (AdvanSure) assays using the same bronchial washing specimens were retrospectively reviewed. Results: Of the 1,297 patients included in this study, 205 (15.8%) were diagnosed with pulmonary TB. Using mycobacterial culture as the reference method, sensitivity of the Xpert assay using smear-positive specimens was 97.5%, which was comparable to that of the AdvanSure assay (96.3%, p=0.193). However, the sensitivity of the Xpert assay using smear-negative specimens was 70.6%, which was significantly higher than that of the AdvanSure assay (52.9%, p=0.018). Usng phenotypic drug susceptibility testing as the reference method, sensitivity and specificity for detecting rifampicin resistance were 100% and 99.1%, respectively. Moreover, a median turnaround time of the Xpert assay was 1 day, which was significantly shorter than 3 days of the AdvanSure assay (p<0.001). Conclusion: In comparison with the AdvanSure assay, the Xpert assay had a higher sensitivity using smear-negative specimens, a shorter turnaround time, and could reliably predict rifampin resistance. Therefore, the Xpert assay might be preferentially recommended over TB-PCR in Korean TB diagnostic algorithm.

Screening of Sterol Biosynthesis Inhibitors from Natural Products Using Recombinant Yeast Carrying Human Lanosterol Synthase

  • Sung, Chung-Ki;Kim, Eun-A;Chu, Yun-Ho;Shibuya, Masaaki;Ebizuka, Yutaka
    • Natural Product Sciences
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    • v.9 no.4
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    • pp.299-303
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    • 2003
  • For the screening of inhibitors of sterol biosynthesis from natural products, a simple and rapid assay method was developed using recombinant yeast carrying human lanosterol synthase, main target of this assay method. Sterol biosynthesis inhibition activity was monitored only by the inhibition of growth of the recombinant yeast. By changing the substrate, this assay method can figure out which step is inhibited in the sterol biosynthesis by the test material. With this assay method total 102 plant samples were screened for their inhibitory activity of sterol biosynthesis. Among plant water extracts screened, 11 plant samples showed inhibitory activity on sterol biosynthesis in ergosterol (-) medium. For selection of the specific inhibitory materials, 11 plant samples were reassayed in ergosterol (+) medium. After all 5 plant samples, Abutilon avicennae Gaertn. (stem), Alnus japonica Steud. (stem), Amaranthus mangostanus L. (aerial part), Philadelphus schrenckii Pupr. (leaf) and Pimpinelia brachycarpa Nakai (aerial part), showed specific inhibitory activity.

Optimization of Penicillin Amidase Production and A Simplified Enzyme Assay Method (페니실린 아미다제 생산의 최적 조건 및 간이화한 효소 정량 방법에 대한 연구)

  • 김경훈;유두영
    • Korean Journal of Microbiology
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    • v.15 no.4
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    • pp.145-153
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    • 1977
  • Penicillin amidase (EC 3.5.1.11) was produced by a mutant strain of Bacillius megaterium ATCC 14945. Hydroxylamine assay method for the determination of 6-APT was modified by using "HCl addition techniques" in order to simplify the time consuming orginal assay method without sacrifice of accuracy. Using the new mutant strain, the effects of fermentation conditions on enzyme production were studied.e studied.

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Improved Fluorometric Assay Method for Ribonuclease Activity

  • Lee, Jong-Soo;Choi, Jong-Soo
    • BMB Reports
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    • v.30 no.4
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    • pp.258-261
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    • 1997
  • A simple quantitative assay method for ribonuclease activity has been developed. This method is based on the decrease of fluorescence intensity emitted by the ethidium bromide bound to RNA due to the degradation of RNA by ribonuclease. The substrate RNA was reacted with ribonuclease A and the fluorescence intensity was measured after the addition of ethidium bromide. The intensity difference was calculated using a blank reaction mixture containing no RNase. Whole cellular RNA substrate produced a significant error and was not suitable for this assay method possibly because of local microheterogeniety caused by high molecular weight rRNA. but satisfying results were obtained with tRNA substrate. The intensity difference increased linearly by raising enzyme concentration up to $2{\times}10^{-4}$ Kunitz Units of ribonuclease A. More refined and reliable results were obtained by use of initial reaction velocities which were calculated from the plots of intensity difference vs time. A linear relationship between initial velocities and enzyme concentrations was observed up to 0.01 Kunitz Units of enzyme.

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Use of cccDNA Breakage Assay and Sulforhodamine B Assay for the Prescreening of Antitumor Agents from Microbial Sources (미생물 배양액으로부터 항암제의 예비선별을 위한 cccDNA Breakage 활성검정과 Assay Sulforhodamine B 활성검정의 이용)

  • Lee, Sang-Han;Lee, Dong-Sun;Kim, Jong-Guk;Hong, Soon-Duck
    • Journal of Life Science
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    • v.8 no.1
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    • pp.67-71
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    • 1998
  • In order to develop new antitumor agents from fermentation broths, we used cccDNA breakage assay abd sulforhodamine B assay for prescreening. As a result, it was shown that sample reach 3.3% when using cccDNA breakage assay. In sulforhodamine B assay, we obtained 4 acive fraction against A549 (a cell line of human lung carcinoma) and SK-OV-3 (a cell line of human adenocarcinoma). These results suggest that these assay would be a promising method for antitumor prescreening from microbial sources.

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Antioxidant Activity Analysis of Useful Compounds from Artemisiae Annuae Herba Using On-line Screening HPLC-ABTS+ Assay (On-line Screening HPLC-ABTS+ assay를 이용한 청호로부터 유용성분의 항산화 활성 분석)

  • Lee, Kwang Jin;Ma, Jin Yeul
    • Journal of Applied Biological Chemistry
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    • v.57 no.4
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    • pp.301-305
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    • 2014
  • The Antioxidant activity screening identification of five kind compounds in Artemisiae annuae herba with the on-line screening high performance liquid chromatography (HPLC) $ABTS^+$ assay. The various experimental variables such as the extraction time (h) and extraction solvent composition (%) of dipping method were investigated efficiently extraction at the room temperature $25^{\circ}C$. The results, the highest yield of total extract amount (0.458 g, 15.250%) was obtained by dipping method with 100% water and extraction time to 3 h. And the on-line screening HPLC-$ABTS^+$ assay method was rapid and efficient to search for bioactivity from natural products.

Quantitative Assessment of Xenoestrogenic Environmental Pollutants using E-SCREEN Assay (E-SCREEN Assay를 이용한 내분비계 장애물질의 정량적 평가)

  • 오승민;이상기;정규혁
    • YAKHAK HOEJI
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    • v.44 no.5
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    • pp.416-423
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    • 2000
  • There is a growing concern that a wide variety of chemicals released into the environment can disrupt the endocrine system of fish, wildlife and humans. Endocrine disrupting chemicals (EDCs) include pesticides such as DDT lindane and atrazine, the food packaging chemicals, phthalates and bisphenol A, alkylphenol ethoxylate detergents and the chemical industry by-products, dioxins. Xenoestrogens in the environment have been argued about health risk, because of estrogen mimetic chemicals are exposed only small amounts to human. A number of in vivo and in vitro assays are now in use to assess the activity of xenoestrogens in the environment. A human breast cancer cell line (MCF-7) was used to develop in vitro screening assay for the detection of xenoestrogenic environmental pollutants. The E-SCREEN (MCF7-BUS) assay is proposed as a reliable, easy and rapid-to-perform method. To optimize and validate this method before it can be used routinely, several phenol compounds and pesticides suspected to be estrogenic were tested using I-SCREEN assay. The results showed that this method is a valuable tool for screening potential estrogen-mimicking environmental pollutants and quantitative determination of estrogeniciy.

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Direct Colorimetric Assay of Microcystin Using Protein Phosphatase

  • Oh, Hee-Mock;Lee, Seog-June;Kim, Jee-Hwan;Park, Chan-Sun;Yoon, Byung-Dae
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.6
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    • pp.418-421
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    • 2000
  • A new direct colorimetric assay of microcystin in water and algal samples is proposed consisting of two procedures as follows: 1) the elimination of phosphorus in the sample and concentration of microcystin using a C(sub)18 cartridge, 2) the detection of the released phosphorus by the ascorbic acid method and determination of protein phosphatase (PP) inhibition by microcystin. The optimum amounts of phosphorylase ${\alpha}$ and PP-1 in 50 ${\mu}$L concentrated sample were 50$\mu\textrm{g}$/50${\mu}$L buffer and 1.0unit/50${\mu}$L buffer, respectively, for the best assay. The pH for the maximum activity of PP-1 was 8. The minimum detectable concentration for this method was about 0.02$\mu\textrm{g}$/L, which is sufficient to meet the proposed guideline level of 1$\mu\textrm{g}$ microcystin/L in drinking water. Consequently, it would seem that the proposed direct colorimetric assay using PP is a rapid, easy, and convenient method for the detection of microcystin in water and algal samples.

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