• 제목/요약/키워드: Microdilution assay

검색결과 40건 처리시간 0.03초

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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    • 제21권3호
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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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Evaluation and Optimization of a Serum-based Minimum Inhibitory Concentration Assay to Caspofungin in Candida albicans Clinical Isolates

  • Yoo, Young Bin;Kim, Sung-Soon;Kim, Young Kwon;Kim, Sunghyun
    • 대한의생명과학회지
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    • 제22권4호
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    • pp.174-183
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    • 2016
  • In the present study, a serum-based minimum inhibitory concentration (MIC) testing to caspofungin was optimized and evaluated to solve the limitations of the conventional Clinical and Laboratory Standards Institute (CLSI) guideline-based antifungal agent MIC test and the usefulness of this testing for clinical application was determined. A total of 105 Candida albicans clinical isolates were used for measuring MIC to caspofungin. Results showed that growth characteristics were different according to types of serum and the mouse serum was the most suitable for this assay. In order to measure the optimal concentration of mouse serum, 0 to 100% mouse serum were added to the media during fungal culture. The optimal concentration of serum was 50% when consideration of antifungal agent administration and inoculum size, serum components and ease of hyphae separated, and the consideration of the degree of growth. In comparison of the usefulness between the conventional Alamar-modified broth microdilution MIC assay and 50% mouse serum-based MIC testing, the range of $MIC_{80}$ of the Alamar-modified broth microdilution MIC assay was $0.13{\sim}2.0{\mu}g/mL$ (SD ${\pm}0.42{\mu}g/mL$) and that of the 50% mouse serum-based MIC assay was $2.0{\sim}32.0{\mu}g/mL$ (SD ${\pm}9.01{\mu}g/mL$). The range of $MIC_{50}$ of the Alamar-modified broth microdilution MIC assay was $0.13{\sim}2.0{\mu}g/mL$ (SD ${\pm}0.40{\mu}g/mL$) and that of the 50% mouse serum-based MIC assay was $1.0{\sim}16.0{\mu}g/mL$ (SD ${\pm}2.36{\mu}g/mL$). The MICs of 50% mouse serum-based MIC testing was increased by up to 4 to 64 times than Alamar-modified broth microdilution MIC assay. In conclusion, a 50% mouse serum-based MIC assay was more useful for measuring MIC in Candida albicans clinical isolates than conventional colorimetric broth microdilution MIC testing.

Antifungal Activity of Bee Venom and Sweet Bee Venom against Clinically Isolated Candida albicans

  • Lee, Seung-Bae
    • 대한약침학회지
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    • 제19권1호
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    • pp.45-50
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    • 2016
  • Objectives: The purpose of this study was to investigate the antifungal effect of bee venom (BV) and sweet bee venom (SBV) against Candida albicans (C. albicans) clinical isolates. Methods: In this study, BV and SBV were examined for antifungal activities against the Korean Collection for Type Cultures (KCTC) strain and 10 clinical isolates of C. albicans. The disk diffusion method was used to measure the antifungal activity and minimum inhibitory concentration (MIC) assays were performed by using a broth microdilution method. Also, a killing curve assay was conducted to investigate the kinetics of the anti-fungal action. Results: BV and SBV showed antifungal activity against 10 clinical isolates of C. albicans that were cultured from blood and the vagina by using disk diffusion method. The MIC values obtained for clinical isolates by using the broth microdilution method varied from $62.5{\mu}g/mL$ to $125{\mu}g/mL$ for BV and from $15.63{\mu}g/mL$ to $62.5{\mu}g/mL$ for SBV. In the killing-curve assay, SBV behaved as amphotericin B, which was used as positive control, did. The antifungal efficacy of SBV was much higher than that of BV. Conclusion: BV and SBV showed antifungal activity against C. albicans clinical strains that were isolated from blood and the vagina. Especially, SBV might be a candidate for a new antifungal agent against C. albicans clinical isolates.

In vitro Antifungal Activity of Limonene against Trichophyton rubrum

  • Chee, Hee-Youn;Kim, Hoon;Lee, Min-Hee
    • Mycobiology
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    • 제37권3호
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    • pp.243-246
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    • 2009
  • In this study, the antifungal activities of limonene against Trichophyton rubrum were evaluated via broth microdilution and vapor contact assays. In both assays, limonene was shown to exert a potent antifungal effect against T. rubrum. The volatile vapor of limonene at concentrations above $1{\mu}l$/800 ml air space strongly inhibited the growth of T. rubrum. The MIC value was 0.5% v/v in the broth microdilution assay. The antifungal activity of limonene against T. rubrum was characterized as a fungicidal effect.

Clarithromycin 약제에 대한 Mycobacterium abscessus 임상 내성 균주 분리 (Detection of Clarithromycin-resistant Strains from Clinical Isolates of Mycobacterium abscessus)

  • 이승헌;박영길;류성원;심명섭;류우진;김희진
    • Tuberculosis and Respiratory Diseases
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    • 제64권6호
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    • pp.422-426
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    • 2008
  • 연구배경: Mycobacterium abscessus는 빠른 성장성을 지닌 비결핵균중에서 높은 병원성과 약제 내성을 나타내는 종이며, clarithromycin과 azithromycin 항결핵제가 M. abscessus에 효과가 있는 유일한 경구용 항결핵제이다. 본 연구에서는 역교잡반응법과 약제감수성검사법을 이용하여 clarithromycin 약제에 대한 M. abscessus 임상 내성균주 검출을 시도하였다. 방 법: 역교잡반응법과 약제감수성검사법을 이용하여 220개의 M. abscessus 임상 균주를 대상으로 내성 균주를 분리하였다. 결 과: 약제감수성검사법으로 7개의 임상 내성 균주들을 검출하였고, 이들 중 3개의 내성 균주는 점 돌연변이 균주로서 역교잡반응법으로도 확인하였다. 결 론: M. abscess 균주에서는 점 돌연변이 및 다른 종류의 내성 특성을 나타내고 있음을 확인할 수 있었다.

감마선과 UV 조사에 의한 항생제 분해 (Assays of Residual Antibiotics after Treatment of γ-ray and UV Irradiation)

  • 신지혜;남지현;유승호;이면주;이동훈
    • 방사선산업학회지
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    • 제4권1호
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    • pp.39-45
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    • 2010
  • The pollution of antibiotics is a major cause of spreading antibiotics resistant bacteria in the environment. Applications of ozonation, UV, and ${\gamma}-ray$ irradiations have been introduced to remove antibiotics in the effluents from wastewater treatment system. In this study, we compared the chemical (HPLC) and biological (antimicrobial susceptibility test, AMS) assays in measuring of the concentrations of residual antibiotics after ${\gamma}-ray$ and UV irradiation. Most samples were degraded by ${\gamma}-ray$ irradiation (1~2 kGy). However, lincomycin and tetracycline were not degraded by UV irradiation. The concentration of residual antibiotics, that was treated with ${\gamma}-ray$ and UV irradiation, measuring by bioassay was similar to HPLC. The concentrations of ${\gamma}-ray$ irradiated cephradine measured by AMS test were 2 times higher than that of HPLC assay, indicating AMS test is more sensitive than HPLC assay. These results indicate that ${\gamma}-ray$ irradiation technique is more useful than UV irradiation, and biological assay is more useful to detect the antibiotics and toxic intermediates in antibiotics degradation.

Endophytic Diaporthe sp. ED2 Produces a Novel Anti-Candidal Ketone Derivative

  • Yenn, Tong Woei;Ring, Leong Chean;Nee, Tan Wen;Khairuddean, Melati;Zakaria, Latiffah;Ibrahim, Darah
    • Journal of Microbiology and Biotechnology
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    • 제27권6호
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    • pp.1065-1070
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    • 2017
  • This study aimed to examine the anti-candidal efficacy of a novel ketone derivative isolated from Diaporthe sp. ED2, an endophytic fungus residing in medicinal herb Orthosiphon stamieus Benth. The ethyl acetate extract of the fungal culture was separated by open column and reverse phase high-performance liquid chromatography (HPLC). The eluent at retention time 5.64 min in the HPLC system was the only compound that exhibited anti-candidal activity on Kirby-Bauer assay. The structure of the compound was also elucidated by nuclear magnetic resonance and spectroscopy techniques. The purified anti-candidal compound was obtained as a colorless solid and characterized as 3-hydroxy-5-methoxyhex-5-ene-2,4-dione. On broth microdilution assay, the compound also exhibited fungicidal activity on a clinical strain of Candida albicans at a minimal inhibitory concentration of $3.1{\mu}g/ml$. The killing kinetic analysis also revealed that the compound was fungicidal against C. albicans in a concentration- and time-dependent manner. The compound was heat-stable up to $70^{\circ}C$, but its anti-candidal activity was affected at pH 2.

조릿대 Ethanol 추출물 및 분획물의 항균 효과 (Antibacterial Activity of Ethanol Extract and Fraction of Sasa borealis)

  • 장미란;이다움;김건희
    • 한국식품조리과학회지
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    • 제26권6호
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    • pp.848-852
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    • 2010
  • This study investigated the antibacterial activities of extracts and fractions of Sasa borealis against eight bacteria (Bacillus cereus, Bacillus subtilis, Listeria monocytogenes, Staphylococcus aureus, Psedomonas aerginosa, Salmonella choleraesuis, Serratia marcescens and Vibrio vulnificus) by broth dilution assay. Using survival curves, the kinetics of bacterial inactivation upon exposure to the extracts and fractions were followed for 24 h. In this same manner, MIC (minimum inhibitory concentration) values were determined by broth microdilution assay and then confirmed to be the extract concentrations that inhibited bacterial growth. Sasa borealis extracts showed antibacterial activities against all tested bacteria. In particular, all tested fractions of Sasa borealis had stronger activities than 70% ethanol extract. MIC of Sasa borealis extract was determined to be 5 mg/mL against Salmonella choleraesuis. All fractions of Sasa borealis extract had extremely strong antibacterial activities. MIC of fractions were determined to be 0.03~2.5 mg/mL. These results suggest that the extracts and fractions of Sasa borealis effectively inhibited bacterial growth and thus are useful as natural antibacterial agents.