General Principles for the Cultivation of MAP
The cultivation of MAP is based on traditional methods for the culture of slowly growing mycobacteria from clinical samples (Merkal et al., 1964; Gillespie, 1999). There are four critical steps, which will be described in more detail in the sections that follow:
1.
Decontamination of clinical samples to destroy or suppress irrelevant, mostly rapidly growing microorganisms, which include both bacteria and fungi. These are present in vast numbers in faeces and most types of environmental samples, but there are fewer in tissues, blood or milk.2. Prolonged incubation in an appropriate medium containing antimicrobial agents to suppress any remaining contaminants for long enough for MAP to emerge.
3. Recognition of MAP colonies on a solid medium, or observation of a particular sign of growth or reaching a prescribed incubation period endpoint in a broth medium.
4. Identification of MAP by phenotypic and/or genotypic means.
5. Subculture to confirm phenotype (mycobactin dependence), or for molecular typing.
18.4
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