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Mycobacterium tuberculosis is a slow-growing bacterium that is the etiological agent of tuberculosis. Agar-based and egg-based media incorporating green malachite and Middlebrook broths or solid media are recommended as the “gold standard” for isolation, culture, and definite diagnosis of M. tuberculosis (6).
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Neg., negative result. Under these culture conditions, 58.3% of all mycobacteria were detected within 14 days, 37.5% were detected within 21 days, and 4.2% were detected within 28 days. Fifty percent of species in the M. tuberculosis complex were detected within 14 days, and 50% were detected within 21 days.
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Middlebrook 7H10 agar contains salts, vitamins and malachite green and is the most widely used for mycobacteria.
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Mycobacteria are Gram-positive, catalase positive, non-motile, non-spore forming rod-shaped bacteria (0.2–0.6 μm wide and 1.0–10 μm long). The colony morphology of mycobacteria varies with some species growing as rough or smooth colonies. Colony colour ranges from white to orange or pink (Iivanainen, 1999).
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Mycolic acids are unique long chain fatty acids found in the lipid‐rich cell walls of mycobacteria including the tubercle bacillus M ycobacterium tuberculosis. Essential for viability and virulence, enzymes involved in the biosynthesis of mycolic acids represent novel targets for drug development.
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Lowenstein-Jensen (LJ) medium is most widely used for tuberculosis culture. LJ medium containing glycerol favours the growth of M. tuberculosis while LJ medium without glycerol but containing pyruvate encourages the growth of M.
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tuberculosis is very restricted in growth by acid pH and requires higher concentrations of Mg2+ for growth at a pH of 6.5 or lower. Using a defined medium (Sauton) containing moderate levels of Mg2+ (100 μM), the growth of M. tuberculosis was reduced by a pH of 6.25 and was almost completely absent at pH 6.0.
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The gold standard method for detecting Mycobacterium tuberculosis , which is the aetiological agent of TB, is still based on microbiological culture procedures, followed by species identification and drug sensitivity testing. Sputum is the most commonly obtained clinical specimen from patients with pulmonary TB.
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