Lab Identification Of Mycobacterium Tuberclosis 17.4.2014.
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Transcript of Lab Identification Of Mycobacterium Tuberclosis 17.4.2014.
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Lab Identification Of Mycobacterium Tuberclosis 17.4.2014
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ObjectivesAt the end of the practical demonstration, student should be able to:Identify mycobacteria on the basis of their:• Acid fast staining.• Morphology as seen on microscopy• Culture medium• Differentiate between gram staining and acid fast staining• Clinical features of tuberclosis
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Mycobacterium
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Differences and similarities between Actinomyces,Nocardia and Mycobacteria
GenusGenus SporeSpore Aerobic Aerobic GrowthGrowth
ExotoxinExotoxin Facultative Facultative IntracellularIntracellular
IC IC HostsHosts
Acid Acid FastFast
Branching Branching rodsrods
ActinomycesActinomyces __ __ __ __ __ __ ++
NocardiaNocardia __ ++ __ __ ++ ++ ++
MycobacteriumMycobacterium __ ++ __ ++ ++ ++ __
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Mycobacterium TUBERCLOSIS MORPHOLOGY
•Are slender bacilli exhibit filamentous form resembling fungal mycelium(myces meaning fungus)•Stained by hot carbol-fuchsin and resist declorization by dilute mineral acids this why refered to acid –fast bacilli• NONMOTILLE•NONSPORING•NONCAPSULATED BACILLUS
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Labrotary diagnosis of mycobacteria
1- Detection Microscopy – carbolfuchsin acid fast test (Z.N stain)Morning sputum collection for 3 days Culture-A- solid egg-based LJ(lowenstien jensn medium B- solid agar –based or broth –based Middlebrook medium
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2- Identification •Morphological properties•Biochemical reaction•Analysis of cell wall lipids•Nucleic acid probe 3- Skin test Mantoux test
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IDENTIFICATION OF M. tuberculosis
• • Growth rate slow• • Growth temperature 35°-37°C only• • No pigmentation• • Niacin positive• • Nitrate positive• • Catalase negative at 68°C• • growth on LJ medium
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CULTURE OF MYCOBACTERIA TB•Lowenstien-jensn medium•Middlebrook broth media•Tubercle bacilli are aerobes•Grow slowly12-24 h and rquire 8 weeks for incubation•Enreched media ; containing egg potatoes , serum and meat.•NICAIN +
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MORPHOLOGYMICROSCOPY
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The term acid fastness refers to resistance showed by these bacteria to decolorization by acid alcohol(95%ethyl alcohol and 3% Hcl).At this strength acid alcohol decolorizes all bacteria except for mycobacteria.
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Colonies on LJ medium take 18-24 days to appear .They appear as dry,rough,creamy or buff colored colonies.
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Skin test• Mantoux test•Intradermal test 0.1 ml of PPD•Result : positive reaction local edema after 48-72 h of 10 mm diameter •Reagents• old tuberculin test –koch test•PPD Purified Protien derivative
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Clinical Disease and symptoms•Primary infection •Active tuberclosis(chronic caseous granuloma)•Miliary tuberclosis•Symptoms :•chronic cough - productive cough • fever (sweating)•Weight loss
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Treatment and VacinationComined antituberclous drugs:• Isonazid• Rifampicin• Ethambutol • Streptomycin
Vaccination:• BCG (Live attenuated vaccine)
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Thank you for your attention
Dr.Mashair Abdelrahman