GRAM POSITIVE BACILLI Mycobacterium Tuberculosis Complex (MTBC)
Pitong Capreng May Munting TBACO
M. pinipedi, M. caprae, M. mungi, M. microti, M. tuberculosis, M. bovis, M. africanum, M. canetti, M. orygis
Microorganism Other Name/
Characteristics/Morphology
(Microscopic and Macroscopic)
Mycobacterium KOCH BACILLUS
tuberculosis  Gram (+), GRAM GHOST,
GRAM NEUTRAL
 Obligate aerobe, capnophilic
 Slighty curved, 0.2-0.6 um
diameter; 1-4 um length
 CAULIFLOWER COLONIES
in Lowenstein-Jensen (LJ)
medium
STAYS ALIVE IN:
Droplets in AIR – 8-10 days
Putrifying sputum – weeks
Dried sputum – 6-8 months
KILLED BY:
Direct sunlight
 Culture –2 hrs
 Sputum -20-30 hours
Chemical- 5% phenol
 24 hrs
Moist heat, 10 mins boiling,
pasteurization, autoclave
Virulence Factors Clinical Significance/ Disease Assoc Lab Diagnosis
Biochemical Tests/ Culture
CORD FACTOR TUBERCULOSIS SPX: Sputum sa Grande SB CUP DECONTAMINATION & DIGESTION OF SPUTUM
 cord formation -can be disseminated to other parts of the body  Sputum 1. N-acetyl L-cysteine (NALC)- digesting
 trehalose-6-6- (HM Grande SB)  Heart, meninges, spine, bones,  Gastic aspirate – best spx for infants NaOH (2-4%)- decontamination (BEST)
dimycolate genitourinary tract  Secretion from bronchoscopy
“SERPENTINE  Blood 2. Trisodium phosphate
CORDS” POT’S DISEASE – TB of the spine Benzalkonium chloride (Zephiran)
 CSF- pellicle/weblike clot
 MTB arranged in MILIARY TUBERCULOSIS- disseminated TB
 Urine
parallel chains 3. Dithiothreitol & NaOH
(Middle or lower Lungs) accumulation of macrophages  Peritoneal, Pericardial, Pleural –
 inhibits the fusion of (ADA) Adenosine deaminase (>40:
phagosome and with phagocytosed MTB  giant Langhan cells  ISOLATION, DETECTION & IDENTIFICATION
Langhan cells + fibroid cells  tubercle Tubercular effusion) 1. CULTURE: 8 WEEKS/ 2 MONTHS
lysosome
(granulomatous lesions)  tubercle + lymphadenitis EGG-BASED MEDIUM
METHODS FOR DIAGNOSIS
SULFATIDES GHON focuses  accumulation  GHON complex   Lowenstein-Jensen (LJ)
(calcified) RANKE’S complex 1. Chest Xray
 Petragnani Medium – for highly contaminated
Primary infxn- middle lobe infiltration
LIPIDS specimens (stool used for M. avium-
TX: Secondary infxn- upper lobe/apex
 Mycolic acid- Allows intracellulare complex)
DOTS – Directly Observed Treatment Strategy  infiitration
the intracellular  American Thoracic Society Medium
survival of MTB inside Lasts for 6 months  Dorset Egg Medium
2. Tuberculin Skin Test PPD (Purified
the macrophage Protein Derivative)
 Phospholipids PRIMARY DRUGS USED: RIPES AGAR BASED MEDIA:
-organism is heat killed and precipitated
 First 2 months – RIPE  Duboi’s Oleic Acid Albumin Medium
w/ AMMONIUM SULFATASE
 Last 4 months - RI  Middlebrook 7H10, 7H11
-RESULT: (+) 5-10 mm induration/
 Rifampin- turns body fluids to red-orange redness after 48 hrs  Mitchison’s Medium
 Isoniazid -causes Vit B6 deficiency  PPD- intradermal
(peripheral neuropathy) LIQUID MEDIA:
 Mantoux – intracutaneous
 Pyrazinamide- increased UA crystals   Bactec 12B
 Volmer- patch
Gout  Septi-check
 Von pirquet -scratch
 Ethambutol – Colorblindness to red and  Middlebrook 7H9, 7H12
green
2. DSSM- Direct Sputum Smear
 Streptomycin – not used anymore Microscopy 2. LUCIFERASE REPORTER
Spx acceptance must qualify using the MYCOBACTERIOPHAGE
SECONDARY DRUGS: 3COREK BARTLETTE’S CLASSIFICATION: - detects MTB and DRUG SUSCEPTIBILITY
Ciprofloxacin <10 Squamous epithelial cells, >25 PATTERNS w/in 1-2 days
Cycloserine PMNs  Luciferase is from fireflies
Capreomycin
Ofloxacin 2 sputum samples in one day (1 in the 3. QuantiFERON-TB – MEASURES INTERFERON
Rifabutin early morning and 1 random)- 1 out of 2 is GAMMA LEVELS; not affected in px w/ previous
Ethionamide diagnostic BCG vaccination
Kanamycin Diagnostic rate: 70% Cure rate: 85% Principle: ELISA
Slide with coiled smear – 2x3cm or Spx: Heparinized WB
thumb size
DRUG RESISTANCE Examine under 300 OIO fields before 4. RAPID CULTURE: BACTEC RADIOMETRIC
Monoresistant – 1 drug only reporting as negative CULTURE – MTB UTILIZATION OF
Polyresistant – 2 drugs (but not both R & I) RADIOACTIVE PALMITATE AS CARBON
MDR (Multidrug Resistant)- both R & I SPUTUM ON LOOP IS REMOVED SOURCE. Allows early detection (1-2 wks) – check
XDR (Extensively drug resistant) -both R & I plus USING: for “SERPENTINE CORDS”
secondary drug 1) sand and 70% alcohol
 2) glass beads and 90-95% spirit
Symptoms of TB 3) 5% cresol
COUGH OF 2 WEEKS OR MORE with or without the
following:
 Sputum ACID FAST STAINING (STAINS USED)
 Blood in sputum (Hemoptysis)  ZIEHL NEELSEN- hot method
 Night sweats  KINYOUN – cold method (preferred);
 Weight loss uses tergitol or 10%? phenol
 Fever (low-grade)  FITE FARACO’S-Hematoxylin instead
 Myalgia & Backpain of MB as counterstain for M. leprae
 TRUANTS/ AURAMINE-RHODAMINE
– yellow flurorescence against black
 SPENGLERS- for colobrind (+) black
 PAPPENHEIM- MTB – red, M.
smegmatis - blue
 BAUNGARTENS- MTB- blue, M. leprae
- red
5. GeneXpert – PCR based NAAT – FOR DRUG
RESISTANT MTB (rPob gene)
BIOCHEMICAL TESTS
(+) result for MTB
 Niacin Accumulation test- yellow (rgt.
Cyanogen bromide) VS M. bovis (-)
 Nitrate reduction test (Broth –pink/red;
Strip – blue)
 Pyrazinamidase (vs M. bovis, kansasii)
 MPT64 Antigen
(-) result for MTB
 Tellurite
 T2H susceptibility
 68C/ Heat stable Catalase Test (MTBC)

Mycobacterium TB in animals (cattles)

bovis

M. africanum TB in West and Central Africa Used in BCG vaccine (attenuated M. bovis)

M. canetti TB in East Africa

AFB REPORTING
AFB REPORTING BY NATIONAL STANDARD AFB REPORTING BY CDC
GRADING REPORTING GRADING REPORTING
0 0 per 300 fields - 0 per 300 fields
+n 1-9 per 100 fields +/- Repeated on second 1-2 per 300 fields
1+ 10-99 per 100 fields slide
1+ 1-9 per 100 fields
2+ 1-10 in at least 50 fields
2+ 1-9 per 10 fields
3+ >10 in at least 0 fields
3+ 1-9 per field
4+ >9 per field
National Tuberculosis Association Method
Report number and request another 1-2 per slide
specimen
Rare (1+) 3-9 per slide MIMICS TUBERCULOSIS/ TUBERCULOSIS LIKE INFXN–
Few (2+) >10 per slide NoPaHi
Numerous (3+) >1 per OIF Nocardia asteroides
Paragonimus westermanii
Histoplasma capsulatum

TUBERCULIN SKIN TEST PPD

>5 mm (HONoR)
HIV px TUBERCULOSIS SYMPTOMS
Organ transplant COUGH FOR MORE THAN 2 WEEKS OR MORE
Nodular or fibr in Xray W/ OR WITHOUT
Recent Contact w/ an active TB px  Sputum
 Blood in sputum (Hemoptysis)
>10 mm (HIGH RISK INDIVIDUALS AND LOCATION)  Night sweats
High prevalence countries  Weight loss
IV drug users  Low grade fever esp. afternoon or night
Mycobacteriology lab personnel  Myalgia & backpain
Infants, Children, Adolescents
Comorbid condition

>15 mm Persons w/ no known risk factors

GRAM POSITIVE BACILLI Non-tuberculous Mycobacteria (NTMs)
RUNYON’S CLASSIFICATION
GROUP 1: PHOTOCHROMOGENS
GROUP 2: SCOTOCHROMOGENS
GROUP 3:
NONPHOTOCHROMOGENS
GROUP 4: RAPID GROWERS
NON-CULTIVATABLE (ON AGAR)
M. leprae & M. lepromatosis
BIOCHEMICAL TESTS
Forms pigment in light after being grown in the dark (grows after 7 days)- SLOW GROWING  Heat stable catalase (68C): Rgt: 30% H2O2
MASK (+) M. fortuitum, M. gordonae
M. marinum –causes swimming pool granuloma
M. asiaticum  Tween 80 Hydrolysis: (+) M. kansasii, (-) M. avium intracellulare
M. simiae
M. kansasii – Yellow bacillus (TWEEN 80 Hydrolysis (+))  Arylsulfatase (+) pink red (M. fortuitum- chelonei)
Forms pigment in light or dark (grows longer than 7 days) – SLOW GROWING  Iron uptake (+) growth in 20% ferric citrate (+) M. fortuitum (-) M.
SSX GFT chelonae
M. scofulaceum – causes SCROFULA/ cervical lymphadenitis
M. szulgai - Photochromogen (@ 25C) & Scotochromogen  Urease – (+) M. bovis, M. scrofulaceum, M, gastri
M. xenopi – Scotochromogen & Nonphotochromogen
M. gordonae – Tap Water bacillus  Growth in 5% NaCl –(+) M. triviale
M. flavescens
M. themoresistible
Doesn’t form any pigment (grows longer than 7 days) – SLOW GROWING
MUAHIT
M. malmoense
M. ulcerans – Buruli ulcer, inert bacillus
M. avium- TB in bird/ chicken; Lady Windemere Syndrome
M. haemophilum- beta hemolytic
M. avium-intracelllulare complex- Battey bacillus, prevalent in AIDs px
M. terrae triviale – Radish bacillus
Grows in less than 7 days
FASCPh
M. fortuitium
M. abscessus- SEVERE CHRONIC PULMONARY INFECTION
M. smagmetis
M. chelonei
M. phlei- Hay bacillus
M. leprae OTHER MYCOBACTERIA:
Has TROPISM to PERIPHERAL NERVES M. genavensi- Disseminated infections in AIDS, BACTEC (+)
CULTURE: FOOT PADS of MICE/ ARMADILLO M. paratuberculosis – Crohn’s disease
RXN: HYDROLYZE DIHYDROXY-PHENYLALANINE (DOPA) M. gastri – J bacillus
TEST: LEPROMINE
 Fernandez (Early reaction) – 1-2 days
 Mitsuda (Late reaction)– 3-4 weeks
SPX: TISSUE JUICE  EARLOBE, NASAL SCRAPINGS
TX: Sulfone Dapsone
HANSEN’S DISEASE OR LEPROSY
 Lepromatous – Lepromine (-) CMI (-) Many AFB, Multibacillary, “Leonine Face”
 Tuberculoid- Lepromine (+) CMI (+), Few AFB, Paubacillary