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A Method for Antibiotic Susceptibility Testing: Applicable and Accurate


Ramezan Ali Ataee 1 , Ali Mehrabi Tavana 1 , Seyed Mohammad Javad Hosseini 2 , * , Khadijeh Moridi 1 , Mahdi Ghorbananli Zadegan 1

1 Therapeutic Microbial Toxin Research Center, Baqiyatallah University of Medical Sciences, Tehran, IR Iran, and Department of Medical Microbiology, Faculty of Medicine, Baqiyatallah University of Medical Sciences, IR Iran

2 Management Health Research Center, Baqiyatallah University of Medical Sciences, Tehran, IR Iran, and Molecular Biology Research Center, Baqiyatallah University of Medical Sciences, Mhosseini@bmsu.ac.ir, IR Iran

How to Cite: Ataee R, Tavana A, Hosseini S M, Moridi K, Zadegan M. A Method for Antibiotic Susceptibility Testing: Applicable and Accurate, Jundishapur J Microbiol. Online ahead of Print ; 5(1):341-345. doi: 10.5812/kowsar.20083645.2374.


Jundishapur Journal of Microbiology: 5 (1); 341-345
Article Type: Research Article
Received: January 20, 2011
Accepted: July 1, 2011




Background: Emerging antibiotic resistance in pathogenic bacteria has driven the development of new assays for routine antibiotic testing.
Objectives: The purpose of this study was to evaluate the effects of different organic solvents in preparing two-fold decreases in serial penicillin concentration coated onto 96-well plates to design a method for antibiotic susceptibility testing.
Materials and Methods: Benzyl penicillin was dissolved in each solvent (sterile distilled water, PBS, diethyl alcohol, ethanol, butanol, chloroform, 2-propanol, and acetonitrile). Serial dilutions of each solution were loaded onto a 96-well microtiter plate and incubated at 37°C for 12 h. Next, 200 μL of sterilized Mueller-Hinton broth was added along with 50 μL of bacterial suspension at an adjusted concentration equivalent to 0.5 McFarland standards. The prepared plates were incubated at 37ºC for 24 h. Optical density (OD) was measured at 540 nm.
Results: When comparing the ODs of each sample in 96-well microtiter plates with positive and negative controls, significant antibacterial activity was observed. Most activities ranged from 50 to 200 units of penicillin in samples that were diluted with distilled water, PBS, or isobutyl alcohol as a solvent. Analysis of the results suggested that, when using the aforementioned solvents, the minimum inhibitory concentration of penicillin against a sensitive strain of Staphylococcus aureus was ≥50 units of penicillin.
Conclusions: The results revealed that the accuracy and feasibility of this method can greatly reduce the waiting period of antibacterial sensitivity tests. Additionally, this method is lowcost and could benefit patients who urgently require proper antibiotic therapy.

  • Implication for health policy/practice/research/medical education:
    Applying of this antibiotic sensitivity assay in the diagnostic microbiology laboratories is important. Because, precise determination of MIC and minimum bactericidal concentration can be a valuable guide for physicians and patients will be treated successfully.
  • Please cite this paper as:
    Ataee RA, Mehrabi-Tavana A, Hosseini SMJ, Moridi K, Ghorbananli Zadegan M. A Method for Antibiotic Susceptibility Testing: Applicable and Accurate. Jundishapur J Microbiol. 2012; 5(1):341-5.
  • DOI: 10.5812/kowsar.20083645.2374

© 2012, AJUMS. Published by Kowsar M.P.Co. All rights reserved.


Antibiotic Dilution Optical Density

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