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Protocol:
Protocol 08
Identification of NTM by using PNB media
Principle
: Growth of the Mycobacterium tuberculosis complex (MTC) is
inhibited by p-nitrobenzoic acid (PNB), whereas non-tuberculous mycobacteria (NTM)
are resistant.
In this protocol, PNB added culture medium is used to evaluate its usefulness in the
screening of mycobacteria isolates.
Materials
1. PNB reagent (Dissolve 2.55 gr PNB in 100 ml DW. Filter through a 0.2 mm
membrane filter for decontamination. Add 100 microliter PNB into 4 ml MGIT liquid
medium + 0.5 ml OADC + 0.5 ml sample and the final concentration become 500
ug/ml in 5.1 total volume.
2. MGIT liquid medium
3. OADC
Equipment:
1. 37oC incubator
2. 1.5 ml appendorf tube with a screw cap
3. Microcentrifuge
4. Vortex
5. Waterbath
6. -20oC freezer
Microorganisms
1. M tuberculosis complex
2. M avium
3. M kansasii
Version : 3
Last Change : October 28, 2009
Page 1 of 2
Version : 3
Last Change : October 28, 2009
Page 2 of 2