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For a laboratory exercise of antibiotic susceptibility test in a disk-diffusion method. What are the concentrations of antibiotic applied in the test?

For a laboratory exercise of antibiotic susceptibility test in a disk-diffusion method. What are the concentrations of antibiotic applied in the test?

Basic Clinical Laboratory Techniques 6E
Basic Clinical Laboratory Techniques 6E
6th Edition
ISBN:9781133893943
Author:ESTRIDGE
Publisher:ESTRIDGE
Chapter7: Basic Clinical Microbiology
Section7.2: Bacteriology Specimen Collection And Processing
Problem 3.1CS
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For a laboratory exercise of antibiotic susceptibility test in a disk-diffusion method. What are the concentrations of antibiotic applied in the test?
Materials: Broth cultures of Staphylococcus aureus and Escherichia coli • Mueller-Hinton agar plates . Antibiotics (Ampicillin & Tetracycline) • Sterile Filter Paper disks • Sterile Cotton Swabs • Forceps Procedure: 1. Label two agar plates with group number, date, name of the organism used along the edge of the plate. Divide the bottom of the plates into two sections and label each section with the name of antibiotic used (AM- Ampicillin; Te Tetracycline). Duplicates for each type of bacteria. 2. Dip a sterile cotton swab in the bacterial suspension being tested. Squeeze the swab against the inner wall of the tube to remove excess liquid. 3. Swab the entire plate from top to bottom, edge-to-edge leaving no gaps. Rotate the plate and using the same swab, again swab the entire plate from top to bottom. Allow the plates to dry for five minutes. 4. Sterilize the forceps by placing in alcohol. Flame the forceps to remove the alcohol. (Once they have been sterilized, do not touch the forceps' tips to anything. If the tips touch anything, re-sterilize them.) 5. Use the sterile forceps, pick up a filter paper disk and place on the empty petri dish. 6. Add 20 µL of antibiotic solution being tested on the center of the paper disk. 7. Using the sterile forceps, carefully place the moist disk in the center of the appropriate section of the plate inoculated with S. aureus. Gently press the paper disk onto the surface of agar. 8. Repeat steps 4-6 with other antibiotics. 9. When you have placed antibiotic paper disks on all sections of the plates, invert them and incubate the plates at 37°C for 24 10. Repeat steps 2-8 with E. coli. 11. After incubation, measure the diameter of the zone of inhibition (mm). Examine the plates carefully for well-developed colonies within the zone of inhibition. 12. Compare the susceptibility of bacterial cultures to antibiotics tested. apap EDED Group Ane Date Figure 5.1 Antibiotic susceptibility test.
Transcribed Image Text:Materials: Broth cultures of Staphylococcus aureus and Escherichia coli • Mueller-Hinton agar plates . Antibiotics (Ampicillin & Tetracycline) • Sterile Filter Paper disks • Sterile Cotton Swabs • Forceps Procedure: 1. Label two agar plates with group number, date, name of the organism used along the edge of the plate. Divide the bottom of the plates into two sections and label each section with the name of antibiotic used (AM- Ampicillin; Te Tetracycline). Duplicates for each type of bacteria. 2. Dip a sterile cotton swab in the bacterial suspension being tested. Squeeze the swab against the inner wall of the tube to remove excess liquid. 3. Swab the entire plate from top to bottom, edge-to-edge leaving no gaps. Rotate the plate and using the same swab, again swab the entire plate from top to bottom. Allow the plates to dry for five minutes. 4. Sterilize the forceps by placing in alcohol. Flame the forceps to remove the alcohol. (Once they have been sterilized, do not touch the forceps' tips to anything. If the tips touch anything, re-sterilize them.) 5. Use the sterile forceps, pick up a filter paper disk and place on the empty petri dish. 6. Add 20 µL of antibiotic solution being tested on the center of the paper disk. 7. Using the sterile forceps, carefully place the moist disk in the center of the appropriate section of the plate inoculated with S. aureus. Gently press the paper disk onto the surface of agar. 8. Repeat steps 4-6 with other antibiotics. 9. When you have placed antibiotic paper disks on all sections of the plates, invert them and incubate the plates at 37°C for 24 10. Repeat steps 2-8 with E. coli. 11. After incubation, measure the diameter of the zone of inhibition (mm). Examine the plates carefully for well-developed colonies within the zone of inhibition. 12. Compare the susceptibility of bacterial cultures to antibiotics tested. apap EDED Group Ane Date Figure 5.1 Antibiotic susceptibility test.
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