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coli
Methodologies
E-test by AB-biodisk A dilution test based on the diffusion principle No international recognized interpretation guidelines Expensive for routine usage Not approved by CRL AR Disk diffusion CLSI interpretation guidelines for only two antimicrobials Limited interpretation only detect resistance Not approved in the monitoring by the Commission
Methodologies
Agar dilution Approved for four antimicrobials according to CLSI and 12 according to eucast Laborious and time consuming Broth dilution (microbroth dilution by Trek, VetMic etc.) The Golden Standard Approved for four antimicrobials according to CLSI and 12 according to eucast Expensive but easy to use.
Disk diffusion
Testing condition according to CLSI M45-A Cation-adjusted Mller Hinton agar supp. with 5% sheep blood (Depth 4mm, works also with other types of blood) Direct colony suspension, equivalent to MacFarland 0.5 Incubation temp / time: 36-37C for 48h OR 42C for 24h (not less than 36C and not above 42C) Incubation atm: microaerobic equal to 10% CO2, 5% O2 and 85%N2
E-test
Cation-adjusted Mller Hinton agar supp. with 5% sheep blood works also with other types of blood Direct colony suspension, equivalent to MacFarland 1 Incubation temp / time: 36-37C for 48h OR 42C for 24h (not less than 36C and not above 42C) Incubation atm: microaerobic equal to 10% CO2, 5% O2 and 85%N2 No recommended antimicrobial susceptibility interpretive criteria. According to AB biodisk; resistant isolates can be identified based on the CLSI interpretive criteria for Enterobacteriaceae
Agar dilution
Agar and plate preparation according to CLSI M7-A7 / M100-S18
21
20
22
11
18
17
25 13
16 15
14
21
21
21
20
20
20
22
22
22
11
11
11
18
18
18
17
25 13
17
25 13
17
25 13
16 15
14
16 15
14
16 15
14
Control
0,25
0,5
Micro-broth dilution
Testing condition according to CLSI M7-A7 Cation-adjusted Mller Hinton broth supp. with 5% sheep blood Remember to adjust the content of cations and lyse the blood (also horse and cattle blood works fine) Prepare the inoculum by direct colony suspension, equivalent to MacFarland 0.5
Quality Control
Monitor quality of saline, broth, lysed blood, Macfarland st., temp. and atm. Check the purity of the test suspension for each isolate Use the reference strain C. jejuni ATCC 33560
E-test
Cation-adjusted Mller Hinton agar supp. with 5% sheep
Thank you!
Rene Hendriksen (rshe@food.dtu.dk) www.antimicrobialresistance.dk