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258 Cards in this Set
- Front
- Back
Name the test and what it is for
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Catalase test
Differentiate: - Staphylococcus (+) from Streptococcus (-) - Among small non-branching, non-spore forming aerobic gram positive bacilli: ( + ) Listeria spp. Corynebacterium spp. ( - ) Erysipelothrix rhusiopathiae (H2S+), Arcanobacterium haemolyticum |
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Name the test and what it is for
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Coagulase tube test: detects free coagulase producing organisms
Coagulase-positive staphylococci: Staphylococcus aureus subsp. anaerobius S. a. aureus S. a. delphini S. hyicus S. intermedius (dog bite) S. lutrae Staphylococcus schleiferi subsp. coagulans Coagulase-negative staphylococci: S. saprophyticus S.cohnii subsp. cohnii S. cohnii subsp. urealyticum S. captitus subsp. captitus S. warneri S.hominis S.epidermidis S. caprae S. lugdunensis (+ with bound/slide coagulase test and PYR+) Coagulase test = Rabbit plasma + organism incubated at 35*C and checked at 4 hr and 24 hr |
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Name the test and what it is for
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Coagulase slide test: detects bound (clumping factor) coagulase producing organisms
S. aureus group and S. lugdunensis ( - coagulase tube test) |
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Name the test and what it is for
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Coagulase latex agglutination test: detects bound (clumping factor) coagulase producing organisms
S. aureus group and S. lugdunensis ( - coagulase tube test) |
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Name the test
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Kirby-Bauer disk diffusion antibiotic sensitivity testing showing nosocomial MRSA
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Name the test
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Kirby-Bauer disk diffusion antibiotic sensitivity testing showing community acquired MRSA with positive D-test
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Novobiocin disk/gram stain, name the organism on the right side
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Staphylococcus saprophyticus
Coagulase - Staph. involved in approximately 10-20% of UTI (young females) |
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Name the probable organism
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Clostridium perfringens showing double zone of Beta hemolysis on SBA
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Lecithin Lactose Agar
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Clostridium perfringens precipitates lecithin around colonies (opaque halo)
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thioglycolate broth
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Clostridium perfringens in thioglycolate broth, showing gas production
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Name organism
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Pseudomonas aeruginosa on BAP showing metallic haze colonies
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Mac Conkey agar
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Pseudomonas aeruginosa on MAC
Glucose non-fermenter, oxidase positive |
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KIA/TSI tube, organism?
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K/K:
non-glucose, non-lactose, non-sucrose fermenter No H2S Pseudomonas |
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Type of hemolysis, possible organisms
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Beta-hemolysis
Groups A (pyogenes) C (S. dysgalactiae Subs. equisimilis) G (S. equi Subs. zooepidermicus) B (pyogenes) β-hemolytic enterococci |
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Bacitracin test
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Group A Streptococcus (pyogenes) is susceptible to Bacitracin
Bacitracin resistant Beta-Hemolytics S. milleri group (F) S. dysgalactiae Subs. equisimilis (C) S. equi Subs. zooepidermicus (G) β-hemolytic enterococci |
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PYR test
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Positive in Streptococcus pyogenes (GAS)
AND Beta-hemolytic Enterococci |
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Lancefield testing caveat
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Does not distinguish between large-colonyforming,
pyogenic streptococci of groups A, C and G and small-colony-forming group A, C and G strains of the anginosus or “S. milleri” group |
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This bacterium grew in Regan-Lowe medium forming round, domed, mercury-silver, colored, shiny colonies
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Bordetella pertussis
Culture sensitivity: • If ≥14 days from onset of cough, sensitivity is lower, so serology is sent instead of culture • Caveat: Serology is uninterpretable in immunized or unimmunized patients <11 years of age – These patients are cultured regardless of duration of symptoms |
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Name organism
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Corynebacterium diphtheriae
Non-motile Catalase + |
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Name organism and medium
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Tellurite-containing medium, black colonies of Corynebacterium diphtheriae
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Optochin Susceptibility Test, name organism
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Streptococcus pneumoniae
Optichin sensitive, > 14 mm. inhibition |
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BHI agar, what is preferable for this organisms?
Name organisms: Green arrow Red arrow Blue arrow |
Preferable medium for Haemophilus spp.:
Chocolate agar or Horse blood agar with free X (hemin) and V (NAD) Red (X and V) H. influenzae H. haemolyticus Green (V only) - ("The P's like the V") H. parainfluenzae H. paraheamolyticus Blue (X only) - ("X-Rated") H. ducreyi |
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S. aureus streak
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Haemophilus spp. satellitism
S. aureus produces V and if Beta-hemolytic releases X from SBA |
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Gram + methylene blue: name organism
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Corynebacterium diphteriae
Metachromatic granules with methylene blue |
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BYCE medium, name organism
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Legionella pneumophila
Buffered Charcoal Yeast Extract agar Legionella won't stain with gram on direct specimens, it will from subculture specimens |
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Facultative anaerobes, Acid Fast negative, organism group?
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Actinomyces spp.
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Molar tooth colonies
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Actinomyces israelii
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Acid fast stain
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Nocardia spp
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Chalky-white colonies when young then yellow/orange colonies
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Nocardia asteroides
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Medusa-head colonies
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Bacillus anthracis
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Name organism
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Bacillus anthracis
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Name organism
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Bacillus anthracis
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Optichin disks
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Resistant
PYR positive, Bile esculin positive: Enterococcus spp. PYR negative: Bile esculin positive: S. bovis (D) Bile esculin negative: Viridans |
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Enterococcus spp
Ampicillin Resistant Vanco Sensitive |
Enterococcus faecium non-VRE
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Enterococcus spp
Ampicillin Vancomycin Resistant |
Enterococcus faecium VRE
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Enterococcus spp
Ampicillin, Vancomycin susceptible |
Enterococcus faecalis
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Enterococcus spp
Quad plate |
Test for synergy of aminoglycosides with vancomycin for Enterococci causing bacterial endocarditis
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Urease test, gram and tissue, name organism
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Brucella melitensis
Castaneda biphasic blood culture held for 21 days historically used for culture |
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Common-antigen test (glutamate
dehydrogenase or GDH test) |
Used as a screening test for C. diff. to decide which
specimens to evaluate using another assay (e.g., cytotoxicity assay or toxin EIA) |
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Glucose fermenting
Nitrate to nitrite Oxidase negative Possible organisms with this colony type in Mac Conkey |
Lactose fermenting
E. coli (indole+) Enterobacter spp. (VP test +, catalase+, citrate+) Klebsiella spp. (VP test +, non-motile, mucoid colonies) |
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Hektoen enteric agar
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E. coli
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Hektoen enteric agar
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Salmonella spp.
H2S production |
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KIA-TSI
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E. coli
Enterobacter Klebsiella Acid Slant/Acid Butt (A/A) Shigella Vibrio Alkaline Slant/Acid Butt (K/A) Salmonella Citrobacter Proteus Alkaline Slant/Acid Butt (K/A) + H2S Pseudomonas Alkaline Slant/Alkaline Butt (K/K) |
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CIN agar
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Yersinia enterocolitica
•Septicemia in iron overload syndromes •Mesenteric adenitis – RLQ pain mimics appendicitis •Grows well at 4*C (like Listeria) •CIN agar (Cefsulodin-irgasan-novobiocin) |
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Safety pin morphology
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Yersinia pestis
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TBCS agar
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Vibrio cholerae
Oxidase positive/glucose fermenter KIA: K/A TCBS (Sucrose) Yellow: V. cholerae Green: V. vulnificus, V. parahaemolyticus |
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Gram, identify organism group
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Vibrio spp
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Special BAP medium, at 42°C and @ 5% O2, 10% CO2, 85% nitrogen
• Catalase positive • Oxidase positive |
Campylobacter spp
• Hippurate hydrolysis test – If positive, identifies isolate as C. jejuni – Of Campy species, only jejuni is positive |
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Name organism
Motile |
E. coli
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MUG test positive (β-glucuronidase activity)
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E. coli
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Name organism
Non-motile VP+ |
Klebsiella spp
Indole: + K. pneumoniae - K. oxytoca |
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Name organism
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Proteus spp
Indole: + P. vulgaris - P. mirabilis |
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Name organism
Coagulase - Novobiocin resistant |
Staphylococcus saprophyticus
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Name organism
Thayer Martin Glucose fermenting |
Neisseria gonorrheae
Molecular amplification methods – Permit concurrent detection of CT in single specimen – Allow use of urine as a specimen (non-invasive) – Do not require viable organisms (transport not an issue) – Highly sensitive – Rapid • Disadvantages: – GC DNA may be present up to 3 weeks after successful treatment, so NAATs should not be used as test of cure – Isolates not available for susceptibility testing if treatment difficulties arise – Results inadmissible evidence in medicolegal cases |
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Blue
Pink Yellow Green Magenta |
Blue: GBS
Yellow: Listeria monocytogenes Pink: Pneumococcus Green: Neisseria meningitidis Magenta: H. influenzae |
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Name organism
Thayer-Martin Ferments maltose |
Neisseria meningitidis
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Lazy-Beta hemolysis
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Streptococcus agalactiae
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Name organism
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Streptococcus agalactiae
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Name organism
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Listeria spp.
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Name organism
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Listeria monocytogenes
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Hyperdiploidy (> 52 chromosomes)
t(12:21)/TEL-AML1 - cryptic, use FISH |
Favorable cytogenetics for ALL
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t(9;22) (q34;q11) ABL/BCR (p190)
t(4;11)(q21;23) MLL-AF4 11q23/MLL rearrangments t(1;19)(q23;p13) PBX/E2A Hypoploidy < 40 chromosomes Near tetraploid |
Unfavorable cytogenetics for ALL
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t(4;11)(q21;q23) in ALL
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Expression of CD15 and loss of CD10 (5% of B-ALLs)
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Inv(16) or t(16;16)(p13;q22)
t(8;21)(q22;q22) AML1/ETO |
Favorable cytogenetics for AML
Inv(16) or t(16;16)(p13;q22) Core binding factor β and smooth muscle myosin heavy chain - 10-12% of AML (often younger pts.) - Myeloid sarcoma may be present at diagnosis - Myelomonocytic morphology (M4 Eo) with abnormal eosinophils (basophilic granules) - Auer rods in some cases - AT LEAST 3% of blasts positive for MPO - Blasts often at the 20% threshold - May coexpress CD2 t(8;21)(q22;q22) AML1/ETO (core binding factor) - May present as granulocytic sarcoma - Large blasts with abundant cytoplasm - Pseudo-Chediak-Higashi granules, Auer rods - “AML with maturation” (may have mono diff) - Aberrant coexpression of CD19, maybe CD56 |
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t(15;17)(q22;q21)
+8 t(9;11) (children) t(6;9) |
Intermediate cytogenetics for AML
AML with t(15;17)(q22;q12) - Acute promyelocytic leukemia (typical and microgranular forms) - Associated with DIC; microgranular – high WBC count; typical – low count - Nuclei of blasts often reniform or bilobed - Numerous Auer rods - MPO is VERY strong in both forms (why?) - CD34 and HLA-DR often negative; CD33 bright, may have CD2 coexpression - Response to ATRA and induction chemo good except in t(11;17)(q23;q21) and t(5;17) |
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- 7
- 5 del 7q 11q23 MLL rearrangements inv(3q) Complex abnormalities |
Bad cytogenetics for AML
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MPO in myeloid blasts
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MPO in myeloid blasts
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PAS in erythroid blasts
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"Apple core" nuclei of APML
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APML in BM
• t(15;17) – RARα and PML – All-trans retinoic acid (ATRA) responsive – t(11;17) and t(5;17) – less responsive |
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PAS in ALL
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Normal karyotype
-Y isolated 5q- |
Good cytogenetics for MDS
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20q-
+8 |
Intermediate cytogenetics for MDS
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Complex karyotype
5q- associated with any other abnormality 17p abnormality loss of 7 |
Bad cytogenetics for MDS
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Non-specific esterase (NSE)
ANB (alpha napthyl butyrate) |
Diffuse cytoplasmic positivity in monoblasts
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ANA (alpha napthyl acetate)
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Similar reactivity in monoblasts, but inhibited by sodium fluoride (NaF) (vs. megs, erythroid)
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Hematogones on flow cytometry
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Extra Philadelphia chromosome
trisomy 8 isochromosome 17q trisomy 19 |
CML Clonal progression
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Rouleaux with background staining
high-protein states, reactive or clonal |
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Agglutination (100 oil)
cold agglutinin disease |
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Dual population (40 dry).
This could be seen in partially treated/transfused iron deficiency, sideroblastic anemias, and combined B12/iron deficiency. Transfusion |
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Spherocytes (100 oil).
Spherocytes can be seen in hereditary spherocytosis, autoimmune hemolytic anemia, and acute hemolytic transfusion reaction. Microspherocytes, which are even smaller spherocytes, can be seen in Clostridium perfringens infection and in thermal injury (eg, burns). |
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Oxidative process (100 oil).
Blister cells are smaller red blood cells that lack a central pallor. The blister appears as a vacuole at the red blood cell surface. A thin rim of cytoplasm appears to enclose this vacuole. Bite cells are smaller red blood cells that lack a central pallor and appear to have an oval bite taken out of the red blood cell. Some cells have more than 1 bite per cell. Bite blister, and irregularly contracted red blood cells can be seen in oxidative hemolysis (eg, G6PD deficiency) and unstable hemoglobinopathies. |
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Fragments (100 oil).smaller red blood cells that lack central pallor.
They are jagged in shape and appear as a piece of red blood cells. Note how an occasional spherocyte is seen (indicated by the dotted arrows) and a rare, irregularly contracted RBC (indicated by the thick arrow) is seen. In a microangiopathic (fragmenting) process, occasional spherocytes and irregularly contracted cells are seen; however, this does not represent 2 or 3 separate processes because the fragments are in the majority. |
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Echinocytes (100 oil). Also called burr cells
Renal disease, hypophosphatemia, and blood film artifact. |
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Acanthocytes (100 oil). Also called spur cells
McCleod syndrome Movement disorders Microcytic disorders such as iron deficiency and hemoglobinopathies and in normocytic/macrocytic disorders such as liver disease and hyposplenic states. |
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Eliptocytes (100 oil).
Hereditary eliptocytosis, hemoglobinopathies, and iron deficiency. |
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Hyposplenic changes (100 oil).
The solid arrow is demonstrating a Howell–Jolly body, which is a single, round, smooth, dark blue inclusion body seen in this red blood cell. It represents DNA. The dotted arrow indicates Paphenheimer bodies, which are multiple, jagged and irregular, light blue inclusion bodies seen in this red blood cell. It represents iron deposition. Both these inclusion bodies can be seen in hyposplenic states. |
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Basophilic stippling (100 oil).
Basophilic stippling (RNA) seen in thalassemic states, hemolytic processes, and dysplasia. More coarse basophilic stippling has been described in lead poisoning. |
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Falciparum malaria (100 oil).
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Pelgerized neutrophil (100 oil). The neutrophil on
the left (indicated by the solid arrow) is hypolobulated and has a mononuclear form. This is considered pelgerized. The neutrophil on the right (indicated by the dotted arrow) is normal. It can be seen in myelodysplastic disorders or congential disorders |
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Toxic changes (100 oil). The neutrophils on this
film demonstrate increased dark red––purple granulation, consistent with toxic granulation. |
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Megaloblastic changes (100 oil). A hypersegmented
neutrophil with greater than 6 nuclear segments is seen |
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Various platelet sizes and morphologies (100 oil).
The solid arrow indicates a giant hypogranular platelet. The dotted arrow indicates a giant granular platelet. Giant platelets are larger than the size of a normal red blood cell. |
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Circulating plasma cells (100 oil). Note the rouleaux formation in the red blood cells.
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Cryoglobulinemia (100 oil). Cloud-like pale purple
cryoglobulin deposits are seen in between and overlaying the red blood cells. When the cryoglobulins overlay the red blood cells, they can give the false appearance of irregularly contracted and bite cells, especially if the cryoglobulins are small and not darkly stained. |
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May-Hegglin anomaly, showing Döhle-like inclusions (×1000).
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The Alder-Reilly anomaly may be found in healthy individuals or in those with mucopolysaccharidoses, in which granules are metachromatic (×1000).
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Chédiak-Higashi neutrophils and lymphocytes with large granules
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Inherited Pelger-Huët anomaly (×1000).
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del(13q14.3)
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Good cytogenetics for CLL/SLL
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trisomy 12
del(11q22-23) del(17q)(TP53) |
Bad cytogenetics for CLL/SLL
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Diffuse infiltrate of the BM
Increased proportion of prolymphocytes (between 10% and 55%) CD38 (> 30% of cells) and ZAP-70 Unmutated IgVH gene |
Bad prognostic features SLL/CLL
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MCL
t(11;14)(14q32)(11q13) IGH-CCND1 |
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Blastoid variant/pleomorphic variant
+ 12 + 3q -9q Complex karyotype p53 mutations |
Bad prognostic features for MCL
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MCL in BM: non-paratrabecular infiltrates with histiocytes
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FL t(14;18)(p32)(q21) IGJH-BCL2
Grading: Centroblasts: I: 0-15 per HPF II: 5-15 per HPF IIIa: >15 per HPF IIIb: Sheets of centroblasts |
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FL in PBS
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Paratrabecular deposits of FL in BM
Also seen in T-cell rich B-cell lymphoma |
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- t(11;18) – stomach, lung
– t(1;14) – ocular, parotid, cutaneous – t(3;14) – ocular, thyroid, cutaneous – t(1;14) – lung, small bowel +3 +18 All MZL |
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SLVL
•Moderate quantity of cytoplasm •Polar villi •Nucleoli CD19+, CD20+, sIg+, CD11c+ but unlike HCL: CD103 neg (usually), annexin A1 neg |
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HCL
• Cytopenia: neutropenia, monocytopenia • Immunophenotype – CD19+, CD20+, sIg+, – CD11c+ (very bright), CD25+, CD103+, Annexin A1+ – Bcl‐1+ • TRAP+ |
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BM in HCL:
- Dry tap (increased fibrosis) - Stripped nuclei - Blood lake formation |
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PEL
- HHV-8+ – Like KS and Multicentric Castleman’s • HIV + • Effusion contains large cells with – Immunoblastic, plasmablastic, anaplastic features C t l i li ti HHV8 – Cytoplasmic vacuolization • Neg for B-cell, T-cell, myeloid antigens • Pos for CD45, CD30, CD38, CD138, EMA • Clonal Ig rearrangement + |
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BL
Immunophenotype – CD19+, CD20+, sIg+, CD10+, BCL6+, – CD34 neg, TdT neg, BCL2 neg |
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t(8;14)
t(2;8) t(8;22) – Always involving C-MYC (8) |
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MM
Three prognostic groups – Shortest survival – t(4;14), t(14;16), or del(17p) – Intermediate survival – 13q14 deletions alone – Longest survival – no anomalies or only t(11;14) |
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Essential Thrombocythemia (ET)
• Bimodal, female > male • Most stable MPD • Minimal splenomegaly • Marrow – Hypercellular – Large, hyperlobated megakaryocytes – Clustered, paratrabecular megakaryocytes • Cytogenetics – Abnormal in <10% |
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PF
• Cellular (prefibrotic) phase – Marrow hypercellular with increased megs – Abnormal megs – appearance and location • Fibrotic phase – Leukoerythroblastic pattern – Intrasinusoidal hematopoiesis • Cytogenetics – 50% abnormal: del(20q), del(13q), +8, +9 |
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MPO+SBB:
Granulocytic maturation – M1, M2, M3, M4 N t MPOd d MPO SBB • Note: MPO degrades quickly in wet specimens NSE+NSE NaF Monocytic maturation – M4, M5 • Sodium fluoride (NaF) inhibition specific for monocytic cells |
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APML – microgranular variant
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• Autosomal dominant
• Giant platelets, thrombocytopenia, Döhle-like inclusions • Related to: Fechtner, Sebastian, and Epstein syndromes • Chromosome 22q12-13 (MYH9 gene – myosin) May-Hegglin anomaly |
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• Most cases seen in children,
median age 2 years • History of developmental delay or ophthalmic conditions • Most common: – Neuronal ceroid lipofuscinosis (Batten disease) – Acid maltase deficiency (Pompe disease) – Tay Sachs Vacuolated lymphs |
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Gauchers syndrome
Glucocerebroside deficiency |
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E. vermicularis eggs
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Trichuris ovum
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Fertilized Ascaris egg (L)
Unfertilized Ascaris egg (R) |
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Hookworm egg
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Hookworm rhabditiform larva
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Strongyloides rhabditiform larva
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Capillaria philippinensis ovum
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• All stages seen (early ring forms to gametocytes)
• RBCs enlarged (trophozoites and later stages) • As RBC enlarges, membrane remains smooth • Schuffner’s stippling in intermediate forms • Schizonts: >12 merozoites • Gametocyte fills most or all of host cell • Cycle: 48 hours P. vivax |
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• All stages seen (early ring forms to gametocytes)
• RBCs enlarged (trophozoites and later stages) • As RBC enlarges, membrane shows projections • Schuffner’s stippling in intermediate forms • Schizonts: ≤12 merozoites • Gametocyte does not fill entire host cell • Cycle: 48 hours P. ovale |
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P. falciparum, thick smear
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P. falciparum, thin smear
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P. falciparum, gametocyte
• Heavy parasitemia • Only early ring forms and gametocytes are seen • Few if any schizonts or trophozoites • Rings: double chromatin dots common (“headphones”) • Marginal rings common (applique cells) • RBCs not enlarged • No Schuffner’s stippling • Banana-shaped gametocytes • Cycle: 36-48 hours |
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• Mostly mature forms seen (trophs, schizonts and
gametocytes) • Few ring forms • RBCs not enlarged • No Schuffner’s stippling • Trophozoites can appear as “band forms” • Schizonts: 6−12 merozoites • Cycle: 72 hours P. malariae |
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Babesia, tetrad
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The banana-shaped
gametocyte is diagnostic if seen. • Early rings, no trophs – Applique’ forms – Dual infected cells – Dual chromatin dots P. falciparum |
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• Infected large,
young RBCs – And also causes enlargement – BIG RBCs. • Amoeboid trophozoites • Schuffner’s granules • 12-20 merozoites/ schizont P. vivax |
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• Infects old, small RBCs.
• Heavy, blocky trophs • ‘Band forms • 8-12 merozoites/ schizont • Heavy malarial pigment • ‘Rosette’ forms P. malariae |
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W. bancrofti
Sheathed Vector: Mosquito Nocturnal |
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Loa loa
Sheathed Vector: Mango fly (Chrysops) Diurnal |
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O. volvulus
Unsheathed, from skin, not blood Vector: Black fly (Similium) |
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H. nana egg, iodine wet prep
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Hymenolepis diminuta (rat tapeworm) eggs
• Size: 70-85 μm X 60-80 μm (larger than H. nana) • Smooth shell, thicker than H. nana egg shell • Shell is often yellow to brown in color • Onchosphere (embryo) inside shell • No polar thickenings on embryo membrane • No filaments |
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D. latum egg, iodine wet prep
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Sparganosis
Spirometra spp. |
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S. mansoni egg, saline wet prep
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S. japonicum egg
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S. haematobium egg with hatching miracidium
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• Hermaphrodite worms
• Morphologically similar ova • 130-159 um long • Small operculum, may pop open when cover slip pressed Fasciola & Fasciolopsis |
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Clonorchis/Opisthorchis egg, iodine wet prep
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adult flukes
overall resemble coffee beans, may live 10-20 years • Eggs found in feces or sputum – Prominent operculum – 68-118 um long Paragonimus spp. |
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• Does not form cysts
• Size: 9-12 μm • Trophozoite has 2 nuclei • No observable flagella, but classified as a flagellate • Moves by means of pseudopodia when seen in feces • Fairly strong association between Enterobius vermicularis and D. fragilis infections has been noted—suggesting that D. fragilis may sometimes be transmitted via pinworm eggs Dientamoeba fragilis |
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• Detection of oocysts in stool
• Modified acid fast stain of concentrated stool specimen • Football shaped (oval with tapered ends) • Size: 20-33 μm long X 10-19 μm wide • Single sporoblast inside • Color: Red (MAF stain) Isospora belli |
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Cyclospora and Cryptosporidium oocysts, MAF stain
Detection of oocysts in stool • Modified acid fast stain of concentrated stool specimen • Shape: Spherical • Size: 8-10 μm (twice the size of Cryptosporidium oocysts) • Color: Red (MAF stain) Cyclospora cayetanensis |
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Microsporidial spores in stool, modified trichrome stain
• Histology: PAS, GMS, acid-fast stains highlight organisms in tissue biopsy • Modified trichrome-stained fecal smears can reveal spores • Characteristic orange-red color on modified trichrome stain • Size: 1-2 μm • Shape: round • Red “band” or dot is often present Microsporidia spp |
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Eggs are 60-75 m in
length and 40-50 m wide – Operculum, sometimes difficult to visualize – Small knob on the abopercular end, may or may not be evident; sometimes only a roughening or thickening of the egg wall Diphyllobothrium latum |
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• T. solium: 7-13 uterine branches
• T. saginata: 15-30 branches • Beef tapeworm -- T. saginata – sub-Saharan Africa, the Balkans and Middle East, Latin America, and Russia – 77 million infected • Pork tapeworm -- T. solium – Latin America (especially Mexico), Africa, southeast Asia, and eastern Europe – 10 million infected |
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HSV infected cell monolayer Rounded cells on edge of monolayer Rapid cell killing
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VZV
infected monolayer Foci of sandpaper CPE with rounded cells |
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CMV infected monolayer Focal grape like cluster of rounded cells
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EBV Serodiagnosis
Heterophile antibodies (HA) react with antigens phylogenetically unrelated to the antigenic determinants against which they were raised HA secondary to EBV are detected by their ability to react with horse or cattle rbc’s. (Monospot test) Rise in the first 2 - 3 weeks of EBV infection, then rapidly fall at @ 4 weeks Cannot be used in children < 4 years of age |
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Adenovirus CPE – rounded cells Connected by strands
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CPE of Enterovirus Teardrop and kite like cells
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Classic CPE = Syncytium formation Respiratory syncytial virus CPE
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Ziehl-Neelsen smear from broth culture, M. tuberculosis
Cording factor |
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MTB: Susceptibility Testing
• Rapid method – Broth culture using single “critical concentration” for each drug – Streptomycin, INH, rifampin, ethambutol, PZA • Standard method – Agar proportion – Also uses critical concentrations, but other concentrations sometimes added – Expanded panel of drugs can be tested |
MDR and XDR TB
• MDR TB: TB isolate that is resistant to both isoniazid and rifampin • XDR TB: MDR + resistance to fluoroquinolone and 1 of the 3 injectable drugs (amikacin, kanamycin, capreomycin) |
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, shielded , exposed (R)
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M. kansasii, shielded (L)
M. kansasii, exposed (R) |
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exposed
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• Slow growth
• Temperature preference = 37°C • Thin, long, branching, acid-fast rods on smear from liquid culture • Usually nonchromogenic; some isolates can be pigmented • Identification by DNA probe from culture – All 3 MAC species are recognized by the commercial DNA probe for MAC (Accuprobe by GenProbe, San Diego); does not differentiate between species – Specific probes for M. avium and M. intracellulare exist; MAC-X is negative by both M. avium complex |
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exposed
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• Found in hot water systems—optimum growth
temperature is 42°C • May take 6 weeks or longer to grow because 37°C incubation is not optimal • Causes chronic pulmonary disease in adults with underlying disease (e.g. COPD or bronchiectasis) • Extrapulmonary infection seen only in immunocompromised • Slow growth • Optimum growth at 42°C • Long, thin, branching, acid-fast rods on smear from liquid culture (“atypical morphology”) • Scotochromogenic • Identified biochemically M. xenopi |
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, shielded , exposed (R)
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• Causes cutaneous infection
• Associated with exposure to freshwater fish tanks or saltwater following trauma • “Fish tank granuloma”—single nodular lesion confined to one extremity • Appears 2-3 weeks after inoculation • In US, most common in southern coastal states • Classified as a slow grower, but some strains manifest rapid growth • Primary isolation requires incubation at 30°C • Thin, long, branching, acid-fast rods on smear from liquid culture (“atypical” morphology) • Photochromogenic • Identified biochemically M. marinum |
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• Most commonly recovered non-pathogenic
NTM • Non-pathogenic even in immunocompromised patients • Widely distributed in soil and water • Frequently contaminates respiratory specimens sent for mycobacterial culture M. gordonae |
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Rapid Growers
• All are 3-day arylsulfatase positive • All grow on MacConkey agar lacking crystal violet • Differentiated using 3 biochemical tests: iron uptake, salt tolerance, and nitrate reduction |
M. fortuitum group
• Positive iron uptake • Tolerant to 5% NaCl (heavy growth) • Positive nitrate reductase • Resistant to all standard anti-TB drugs • Many drug options, including amikacin, cefoxitin, quinolones (ciprofloxacin, gatifloxacin, moxifloxacin), clarithromycin, doxycycline, sulfonamides, imipenem) • Drug combinations are recommended M. chelonae: Features • Negative iron uptake • Not tolerant to 5% NaCl (minimal growth) • Negative nitrate reductase • Resistant to all standard anti-TB drugs • Fewer drug options than M. fortuitum – Susceptible to amikacin, imipenem, tobramycin, clarithromycin – Resistant to fluoroquinolones, cefoxitin • Clarithromycin is the drug of choice for localized infections M. abscessus: Features • Negative iron uptake • Tolerant to 5% NaCl (heavy growth) • Negative nitrate reductase • Resistant to all standard anti-TB drugs • Fewer drug options than M. fortuitum – Susceptible to amikacin, cefoxitin, imipenem, clarithromycin – Resistant to fluoroquinolones • Clarithromycin is the drug of choice for localized infections, although resistant strains are emerging |
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Plate morphology
– Cream/white colored, smooth, large colonies – Colonies on BAP show mycelial projections into the agar, called “feet” or “star-like” Candida albicans |
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Microscopic morphology
– In primary smears, relatively large (3-6 μm) budding yeast with pseudohyphae (and sometimes true hyphae) – When grown on morphology medium: • Pseudohyphae with clusters of blastoconidia at the septations • Single terminal chlamydospores (large, thick-walled) • Some true hyphae may be seen as well • Positive by germ-tube test within three hours • Smooth green colonies on Candida Chromagar plate • Detection of β-D-galactoaminidase activity Candida albicans |
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• Plate morphology
– Small colonies, smooth, glistening – White to cream colored Candida glabrata |
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• Microscopic morphology
– In primary smears, small ( 2-5 μm) budding yeast without pseudohyphae or true hyphae – When grown on morphology medium: • Budding blastospores • No other structures • Positive by rapid assimilation of trehalose test (RAT assay) Candida glabrata |
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Antifungal Susceptibility
Candida spp |
• Candida albicans is generally sensitive to azoles and other
antifungals • 10% of Candida glabrata isolates are azole resistant • Another 20-30% have reduced azole susceptibility • C. krusei is intrinsically resistant to fluconazole • C. lusitaniae is considered resistant to amphotericin (despite low MICs in many cases) • C. parapsilosis, C. krusei, C. guilliermondii and C. lusitaniae sometimes exhibit relatively high caspofungin MICs but most are probably susceptible |
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Antigen Detection: Specificity
Cryptococcus neoformans |
• Very few false-positive results, when
performed properly • Cause of false positives: – Trichosporon beigelii infection (cross-reactive polysaccharide is produced) – Contamination with agar and syneresis fluid on agar – Platinum wire loops |
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Antigen Detection: Sensitivity
Cryptococcus neoformans |
• Varies by population, stage or duration of
infection, and assay methodology • In general, highly sensitive (~99%) • At least as sensitive as culture • Most false-negative results are due to extent of infection – Single focal lesions, e.g., pulmonary lesions, may not produce a great quantity of antigen – False negatives are less frequent in disseminated or progressive pulmonary infections, but do occur |
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C. neoformans on BAP
• Positive for urease activity • Positive phenol oxidase test – Detects ability of yeast to produce phenol oxidase on substrates containing caffeic acid – Most frequently used medium is birdseed agar – C. neoformans turns dark brown in 2-5 days; other cryptococcal species do not • Identified biochemically using commercial platforms (e.g. VITEK yeast card) |
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A. fumigatus
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Aspergillus in Tissue (H&E Stain)
Arborescent growth, thin hyphae, 45 degree angle dichotomous branching |
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A. fumigatus
Single row of phialides |
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A. flavus
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A. flavus
Circumferential phialides |
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A. niger
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A. niger
Heavily pigmented |
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A. terreus
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A. terreus
Two rows of phialides |
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Penicillium spp
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Penicillium spp
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Fusarium spp
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Fusarium spp
Canoe-shaped/Banana boat Associated with Bone marrow transplant and corneal infections |
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T. rubrum
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T. rubrum
Trichophyton rubrum Rare Pencil shaped macroconidia Many microconidia |
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Microsporum canis
Rate of growth: moderate; mature within 6-10 days • Colony morphology: Surface is white and fluffy; reverse is lemon yellow to yellow-brown |
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Microscopic morphology: Septate hyphae with
long, spindle-shaped, rough (echinulate) macroconidia which taper to a knob-like end; few if any microconidia • Macroconidia contain >6 cells Microsporum canis |
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Epidermophyton floccosum
• Rate of growth: moderate; mature within 10 days • Colony morphology: surface yellow to brown to olive-gray and velvety; reverse is orange to brownish |
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• Microscopic morphology: septate hyphae, no
microconidia; macroconidia are smooth, clubshaped with rounded ends, and contain 2-6 cells; macroconidia found singly or in characteristic clusters Beaver tail spores – no microconidia Epidermophyton floccosum |
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Zygomycosis (GMS Stain)
In tissue – 90* angle branching, aseptate, ribbon like |
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18 hours (L)
48 hours (R) |
Rhizopus spp
Zygomycetes in general |
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Rhizopus spp
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Mucor
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Absidia spp
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Penicillium marneffei
Red diffusable pigment Uncommon dimorphic fungus Skin lesions in tropics Pneumonia in immune suppressed |
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Scopulariopsis species
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Paecilomyces species
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Trichophyton tonsurans
Epidemic Scalp ring worm Ballooning microconida |
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Microsporum gypseum - soil
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Trichophyton rubrum
Pencil shaped macroconidia Many microconidia |
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Microsporum canis
Dog and cat ringworm White colony/ yellow reverse Tuberculate macroconidia – With very few microconidia |
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Malassezia furfur
Lipophilic yeast – oil required for growth Media used for culture must oil or oil overlay Small budding yeast 2 – 4 μM with collarette |
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Exophila spp, mature and early yeast-like
Black Molds that cause Mycetoma/Chromomycosis/Phaehypho |
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young yeast-like colonies
older fuzzy colonies |
Exophila spp.
Black Molds that cause Mycetoma/Chromomycosis/Phaehypho |
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Cladophialophora bantiana
Black Molds that cause Mycetoma/Chromomycosis/Phaehypho |
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Wangiella species
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Phialophora species
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Alternaria spp.
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Alternaria spp.
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Bipolaris species
Very invasive Skin, nasal sinuses, bone brain |
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Curvularia species
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Scedosporium apiospermum/
Pseudallescheria boydii |
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Exserohilum species
Steroid injections CSF infections |
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Pseudallescheria boydii
• The name “P. boydii” refers to the sexual state of the fungus • P. boydii is the telemorph of Scedosporium apiospermum and Graphium eumorphum (asexual forms) |
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Scedosporium apiospermum
P. boydii Graphium |
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cycloheximide agar (L)
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Scedosporium prolificans,
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Scedosporium prolificans,
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- Grows as a mold at 25°C, grows as a yeast form at 37°C
• Plate morphology: – Slowly growing, fluffy white colonies initially; buff or brown with age – Yellow to yellow-brown reverse • Conversion of the mold to the yeast form at 37°C • Demonstration of specific precipitins by exoantigen testing Histoplasma capsulatum |
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mold form
yeast form |
• Microscopic morphology:
– Large, rounded, single-celled, tuberculate (rough walled) macroconidia on short conidiophores – Pyriform microconidia; may be sessile on the sides of hyphae or on short conidiophores Histoplasma capsulatum |
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Tuberculate [projections]
Macroconidia And Microconidia Histoplasma capsulatum |
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Sepedonium species
Looks like Histoplasma grows in 5 days/monomorphic Large rough spores (7 – 17 um), usually saprophyte No micro-conida are formed |
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H. capsulatum var duboisii
Found in Central Africa Associated with skin and bone lesions 30˚C culture identical to H capsulatum Yeast cells 8 – 10 um in size 2X the size of regular Histoplasma |
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Blastomyces Culture at 37*C
Slow growing @ 4 weeks |
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Culture at 30˚C (L)
Culture at 37*C (R) |
Blastomyces dermatitidis
Culture at 30˚C (L) Pear shaped conidia like lollipops Look alike fungus – Chrysosporium species Do DNA probe to confirm identification Culture at 37*C (R) Yeast cell is 8 – 20 um in size |
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Culture at 30˚C
Requires 2 – 3 days to grow, white waxy / wooly Coccidioides immitis |
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Septated hyphae with chains of thick walled barrel
shaped arthroconidia with dead cells in between – arthroconidia inhaled in nature in culture from media incubated at both 30* and 35*C – No yeast phase Coccidioides immitis Malbranchea species is look alike fungus |
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Thick walled spherules (10 – 80 uM) with
endospores NO YEAST CELLS All stages of development / fragmented spherules Granulomatous inflammation with caseation Coccidioides immitis |
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Rhinosporidium seeberi spherules - > 80 uM
Larger than Coccidioides spherules |
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Culture @ 37˚C
Slow (3- 4 weeks) waxy coral-like yeast Large yeast (10 – 30uM) with multiple daughter buds (2 – 10 uM) Paracoccidioides brasiliensis |
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Young (L)
Old (R) |
MOLD PHASE
30*˚C growth in 3 -5 days Turns brown to black over time Sporothrix schenckii |
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MOLD FORM (30 C)
Septate hyphae with conidia in daisy wheel pattern Sporothrix schenckii |
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YEAST PHASE
At 37˚C small oval yeast cells, elongated 2 – 5 μM, described as cigar bodies Sporothrix schenckii |
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Sporothrix schenckii
Asteroid body known as Splendore-Hoeppli phenomenon can be seen – also seen in: Zygomycetes (mucorales) Aspergillus Blastomycosis Candida |
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Penicillium marneffei
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Brown colonies on Birdseed agar
C. neoformans |
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Cryptococcus gatti
In culture and staining identical to C. neoformans except for L Canavanine glycine bromthymol blue medium – C. gatti = blue C.neoformans = colorless |
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Paroxysmal cold hemoglobinuria, with erythrophagocytosis; the arrow points to a red cell that has been phagocytosed by a neutrophil.
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Macrocytic anemia resulting from congenital dyserythropoietic anemia type 1 also yields a characteristic blood smear, with striking poikilocytosis
CDA type I is transmitted by both parents autosomal recessively and usually results from mutations in the CDAN1 gene. CDA type I is characterized by moderate to severe anemia. It is usually diagnosed in childhood or adolescence, although in some cases, the condition can be detected before birth. Many affected individuals have yellowing of the skin and eyes (jaundice) and an enlarged liver and spleen (hepatosplenomegaly). This condition also causes the body to absorb too much iron, which builds up and can damage tissues and organs. In particular, iron overload can lead to an abnormal heart rhythm (arrhythmia), congestive heart failure, diabetes, and chronic liver disease (cirrhosis). Rarely, people with CDA type I are born with skeletal abnormalities, most often involving the fingers and/or toes |
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Erythrocyte with prominent basophilic stippling (arrow), a result of lead poisoning
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Burkitt’s lymphoma, with three basophilic vacuolated lymphoma cells
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Hypogranular promyelocytic leukemia is shown in Panel B, with two characteristic bilobed leukemic promyelocytes
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Cryoglobulin deposition in a blood sample from a patient with hepatitis C virus infection
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HbC - clams
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HEINZ BODY
require supravital dye - crystal violet (reticulocyte stain) • causes: - denatured/oxidized Hb - G6PD deficiency - unstable hemoglobins • bite cells have had them bitten out |