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Extra info for Antibiotics in laboratory medicine
Agar Depth As noted, agar depth will naturally alter the size of the inhibition zone. Most methods have settled on a depth of 4 mm or a similar amount. This represents a balance between a smaller depth, which is likely to generate a reasonable zone size, and a larger depth, which is designed to reduce the zone size variation due to small variations in depth. A number of studies have demonstrated greater plate-to-plate variation in zone diameter when the agar depth is less than 3 mm (14,44). Time between Inoculation and Disk Placement The period of delay between the inoculation of the agar surface and the placement of the disks prior to incubation will have a significant effect on the ultimate zone diameter.
A further advantage of direct blood culture broth use is that knowledge of the Gram-stain characteristics of the isolate can guide the choice of a test medium. There are some drawbacks to direct testing from blood culture bottles: the presence of high thymidine concentrations in blood culture media will carry over and antagonize the sulfonamides and trimethoprim, and the presence of sodium polyanethol sulfonate (SPS) in older blood culture media to remove antibiotics can antagonize some penicillins, aminoglycosides, and glycopeptides (2).
Clearly, the turbidity should be adjusted subsequently as for the growth method. This method is acceptable in almost all situations, but it slightly more labor intensive. Hence it remains an option. The choice of growth versus direct method depends on laboratory workflow. Almost all standards prefer this method for fastidious organisms such as Haemophilus species and N. gonorrhoeae. Plate Inoculation In almost all circumstances, it is preferable to use a replicator apparatus to inoculate the agar plates.