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Biology
심사에 생체 외에서 국물 Microdilution: 새로운 항진균 성 화합물을 감지 하는 쉽고 빠른 방법
심사에 생체 외에서 국물 Microdilution: 새로운 항진균 성 화합물을 감지 하는 쉽고 빠른 방법
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JoVE Journal Biology
Broth Microdilution In Vitro Screening: An Easy and Fast Method to Detect New Antifungal Compounds

심사에 생체 외에서 국물 Microdilution: 새로운 항진균 성 화합물을 감지 하는 쉽고 빠른 방법

Full Text
21,381 Views
08:54 min
February 14, 2018

DOI: 10.3791/57127-v

Calliandra Maria de-Souza-Silva1, Fernanda Guilhelmelli1, Daniel Zamith-Miranda2,3, Marco Antônio de Oliveira1, Joshua Daniel Nosanchuk2,3, Ildinete Silva-Pereira*1, Patrícia Albuquerque*1,4

1Laboratory of Molecular Biology, Department of Cellular Biology, Institute of Biological Sciences,University of Brasília, 2Department of Microbiology and Immunology,Albert Einstein College of Medicine, 3Division of Infectious Diseases, Department of Medicine,Albert Einstein College of Medicine, 4Faculty of Ceilândia,University of Brasília

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Overview

This article presents an adaptable broth microdilution method for screening antifungal compounds and extracts. The method aims to evaluate the activity of potential antifungal agents using a modified susceptibility test commonly employed in clinical laboratories.

Key Study Components

Area of Science

  • Microbiology
  • Pharmacology
  • Mycology

Background

  • Antifungal therapies are crucial in treating fungal infections.
  • Broth microdilution is a standard method for assessing antifungal susceptibility.
  • Automation can enhance the efficiency of screening processes.
  • Natural and synthetic compounds can be evaluated for their antifungal properties.

Purpose of Study

  • To provide a streamlined method for screening antifungal compounds.
  • To facilitate the initial evaluation of new antifungal agents.
  • To demonstrate the procedure using small volumes and short timeframes.

Methods Used

  • Growing fungal strains for assay preparation.
  • Centrifugation of conical tubes to collect cells.
  • Modification of the broth microdilution susceptibility test.
  • Utilization of automation for efficiency.

Main Results

  • The method allows for rapid screening of antifungal activity.
  • Small volumes reduce reagent costs and waste.
  • Automation increases throughput and reproducibility.
  • Demonstration of the procedure by a postdoctoral fellow enhances credibility.

Conclusions

  • This method is a valuable tool for researchers in antifungal drug development.
  • It offers a practical approach to evaluate the efficacy of new compounds.
  • Future studies can build on this method to explore various antifungal agents.

Frequently Asked Questions

What is the broth microdilution method?
It is a standard laboratory technique used to determine the susceptibility of microorganisms to antimicrobial agents.
How does automation improve the screening process?
Automation increases the speed and consistency of experiments, allowing for higher throughput.
What types of compounds can be screened using this method?
Both natural and synthetic antifungal compounds can be evaluated.
Why is it important to screen antifungal compounds?
Screening helps identify effective treatments for fungal infections, which can be life-threatening.
Who demonstrated the procedure?
Daniel Zamith-Miranda, a postdoctoral fellow, demonstrated the procedure in the article.

항진균 성 화합물 및 추출 물 심사는 간단 하 고 융통성 있는 국물 microdilution 방법입니다.

이 방법의 전반적인 목표는 임상 실험실에서 사용되는 국물 미세 희석 감수성 테스트의 변형을 사용하여 잠재적인 항진균 화합물의 활성을 평가하는 것입니다. 이 방법은 천연 또는 합성 화합물의 곰팡이 활성을 평가하는 데 사용되며, 이는 항진균 요법 개발의 첫 번째 테스트 중 하나입니다. 우리는 특히 실험이 자동화로 수행되는 경우 짧은 시간에 소량을 사용하여 새로운 항진균제에 대한 초기 스크리닝에 대한 지침을 제공합니다.

이 절차를 시연하는 사람은 제 연구실의 박사 후 연구원인 Daniel Zamith-Miranda입니다. 텍스트 프로토콜에 설명된 대로 분석을 위한 곰팡이 균주를 성장시켜 이 절차를 시작합니다. 다음 날에는 실온에서 5분 동안 1200G에서 원추형 튜브를 원심분리하여 세포를 수집합니다.

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