Recall (Nov.

2014)
 ALTERNARIA –FUNGI
 SPECIMEN FOR RUBELLA- BLOOD
 Lysostaphin Resistant- Micrococcus
 GENTAMICIN RESISTANT-ENTEROCOCCUS
BACTERIA
 PRESENT AFTER JAW SURGERY-
VEILLONELLA
 CRITHIDIA LUCILAE- SLE
 MUCOR AND RHIZOPUS- FUNGI PICS
 ACINETOBACTER- NFO, UTI, DRUG
RESISTANT
ALTERNARIA
 basal
body

nucleaus kineoplas Flagellum

t

DS DNA TEST FOR

SLE
Mucor sp. Rhizopus sp.
Recall (Oct 2014)
 N. MENINGITIDIS- VIRULENCE- ENDOTOXIN
 CYSTIC FIBROSIS-BURKHOLDERIA CEPACIA
 (-) CAMP S. agalactiae= Hippurate test, Reverse
CAMP
 3 lactose fermenters, gram neg – E.coli, Kleb,
Enterobacter
 S. aureus, enterococci in stool- normal flora stool
 Citrobacter vs Salmonella- MacConkey, LDC
 Rotavirus EIA positive, next to do? Molecular test
 CD4- inducer
 Osmolarity
ASCP POINTERS- MICRO
 10% sodium hypochlorite= best disinfectant
 Rickettsia test= detects Ab
 Rubella = how to identify its immune class :
test IgM or IgG or both
 Major difference of kinyoun and ZN = process
 Penicillium picture = brush like conidiophore
 Steam under pressure = 121’C, 15 mins
 Dermatophytes = macroconidia and
microconidia
ASCP POINTERS- MICRO
 Lysostaphin Resistant – Micrococcus
 CAMP test is doubtful- do other test (hippurate)

 Do ID & AST- Pathogens (Enterococcus, Staph

not on Lactose fermenters)
 Differentiate Morganella morgani and Prov.
stuartii- Ornithine and citrate
 C. freundi & Sal. typhi – citrate and MacConkey
ASCP POINTERS- MICRO
 Y.enterocolitica and Y. pestis differentiate- Y.
pestis (+) motility and ornithine (+)
 Treponema seen – urethral discharge of male
and female
 Optochin resistant- Strep viridans

 Bloody diarrhea- EHEC

ASCP POINTERS- MICRO
 H. influenza= satellitism = V factor
 Lysine deaminase negative = yellow

 Synergy = 2>1

 E. coli trophozoite motility = pseudopod

 Sal. enteritidis = TSI picture (K/A, gas, H2S)

 Sal. paratyphi A = TSI (K/A no gas, no H2S)

 Aspergillus= immunocompromised diabetic

patient sinuses
ASCP POINTERS- MICRO
 Basitracin test doubtful- do PYR
 Serratia marcescens- Dnase test (+)

 Burkholderia cepacia- cystic fibrosis

 Mucoid colony, Lactose fermenter, indole (+)-

Kleb. oxytoca
 Curvularia – curved organism (picture)

 Alternaria- agent of phaeohyphomycosis (pic)

 Contact lens, corneal infection- Acanthamoeba

CURVULARIA
ALTERNARIA
Syncephalastrum
FUNGI: Opportunistic mycosis:

Penicillium
ASCP-Serology, BB, Immuno
 Amplification of Hep 2 nu cells = PCR,
NASBA, TMA
 Target amplification method- PCR, RT-
PCR,TMA (Transcription mediated
amplification), qPCR- Real time PCR
 Branched chain technology (bDNA)- signal
amplification, hybrid capture assay (HCA),
cleavage based amplification
ASCP-Serology, BB, Immuno
 Probe amplification- ligase chain reaction,
strand displacement amplification (SDA)
 Thermocycler, Taq DNA polymerase, primers-
PCR reagents and materials
 Fc portion of IgG- Rheumatoid factor
 Anti smooth muscle Ab (ASMA)– chronic
active hepatitis
 Inc TSH, Inc thyroid Ab = Graves, hashimoto,
thyroid cancer?
 Rim/peripheral pattern – anti-dsDNA
Rim/peripheral – anti-ds DNA
Speckled=anti- Sm, anti-RNP,
anti- Ro, anti -SSB
Homogenous- ssDNA, histone
ASCP Pointers
 Anti-p24, anti-gp120, anti-gp41 (+) band = report as
reactive in Western Blot
 High IgG, high total protein, low albumin- multiple
myeloma
 CSF IgG oligoclonal band – multiple sclerosis
 Mixed lymphocyte culture – compatible cells; very
low/no blastogenesis
 Low C5- Neisseria infection (N. meningitidis)
 Low C3 – overwhelming infection
 C3b- immune adherence& opsonization
 C5a- chemotaxis and anaphylatoxin
ASCP Pointers
 MHC class I – CD8 cells; class II = CD4
 Anti-EBNA = past EBV ; 3 wks after acute

 Destroyed by enzyme – Duffy &MNSU, Xga

 AHG – detect IgG

 Platelet concentrate from whole blood- 5.5 x

1011 and plateletpheresis= 3 x 1011 platelets
 Leukocyte reduced RBC= 80% RBC ,

WBC 5 x 106
 antiBga, Bgb,Bgc = HLA ag associated
 urine- neutralize anti-Sda (Sid)
 Rabbit erythrocyte stroma – absorbs cold
autoantibodies from plasma
 Anti-IgG (-) & anti-C3d (+) = pre warm tech./
adsorption of cold antibody
 Anti-IgG (+) and anti C3d (+) = elution
 Warm autoantibody= elution
 Best blood group to be transfused to RH positive
infant with RH negative mother = group O negative
 Review Ab panel/screen/ABO discrepancy/ Serial
dilution/ RH
Troubleshooting Incompatible
Crossmatches
Ab Screen AC Major Possible problem
Xmatch

(-) (-) (+) - ABO/Rh typing error

- Donor unit w/ (+) DAT
- Patient w/ low incidence Ab

(+) (-) (+) - Patient alloantibody

(+) (+) (+) - Patient autoantibody

-- Rouleaux
ASCP sample question
 Anti-JKa = delayed hemolytic transfusion rxn
Sample question:
A patient with an anti-K and an anti-JKa in her plasma
needs 2 units of RBA for surgery. How many group
specific units would need to be screened to find 2
units of RBC? The frequency of JKa+ is 77% and K+
frequency is 10%.
A. 6 2 / .207 = 9.66 or 10

B. 10
C. 20
D. 36
A patient with an anti-K and an anti-JKa in her plasma
needs 2 units of RBA for surgery. How many group
specifc units would need to be screened to find 2
units of RBC? The frequency of JKa(-) is 77% and
K(-) frequency is 10%.
A. 10 2/ .077 = 25.97 or 26
B. 20

C. 26
D. 30
Which one has the lowest amount of H
antigen?
A. Type O

B. Type A1
C. Type A2
D. Type B
O negative woman gave birth to an O positive
infant. The fetal blood volume was 5ml. How
many Rh vial(s) should be administered?
A. 1 5/30= 0.16 + 1= 1.16
B. 2
C. 3
D. 4
Which one is an IgG type associated with
PCH?
A. anti-P1

B. Anti-PP1pk

C. Anti-P
D. Anti-pk
The result of a DAT reveals the following:
Anti-IgG (+)
Anti-C3d (+)
What test should be performed next?
A. Antibody screen
B. Antibody test
C. Absorption

D. Elution
The result of a DAT reveals the following:
Anti-IgG (+)
Anti-C3d (+)
Saline control (-)
Polyspecific AHG (+)
What test should be performed?
A. Treat RBC with chloroquine diphosphate
B. Autoabsorption
C. Alloabsorption

D. Elution
The result of a DAT reveals the following:
Anti-IgG (-)
Anti-C3d (+)
Polyspecific AHG (+)
What test should be performed?
A. Treat RBC with chloroquine diphosphate

B. Autoabsorption

C. Alloabsorption

D. Prewarmed technique
ASCP Pointers-CM
 Viralmeningitis – increase lymphocyte
 Bacterial meningitis- increase neutrophil

 Boric acid – preserves protein

 Decrease SG(<1.010) – Diabetes insipidus

 Increase SG (>1.010) – Diabetes mellitus

 constant low SG = renal tubular failure

 Isosthenuria consistent 1.010= failure to

concentrate the urine
ASCP Pointers-CM
 Hemoglobinuria- clear red
 Urine bilirubin – obstructive jaundice

 Urine urobilinogen- hemolytic anemia; Ehrlich’s

 Yellow foam urine- bilirubin (+)

 Synovial fluid diluents

 Sperm ct = process after liquefaction (30-60min)

 Effect of ascorbic acid in glucose test

ASCP Pointers-CM
 Why shift to clinitest if (-) glucose rgt strip
test- detects reducing substance hence more
sensitive
 Glucose reagent strip test- specific

 Uric acid / monosodium urate= birefringent

under polarized light ; negative birefringence
under compensated polarized light
ASCP Pointers- CM
 Cyanide nitroprusside test (+) = cystinuria
 Protein error of indicator= principle in CHON
reagent strip test
 WBC, RBC, Bacteria = hazy urine

 RBC lyzed in alkaline pH

 Yeast mistaken as RBC

 Alkaline urine- protein effect

 (+) nitrite = cystitis, pyelonephritis

ASCP Pointers- chemistry
 Increased chloride in sweat= cystic fibrosis
 Heparin = most blood chemistry analysis

 Hemolysis acceptable= sodium test

 Hemolysis not acceptable = potassium

 Troponin test = Myocardial infarction

 Float, turbid, milky, creamy serum=

chylomicrons
 CA 19-9 and CEA = colorectal cancer
ASCP Pointers- chemistry
 Lp(a)= increased atherosclerotic CAD
 LDL= CAD risk

 Albumin/prealbumin = malnutrition

 Cocaine = benzoylecgonine

 Codeine, Heroine = morphine

 Procainamide = NAPA (N-acetylprocainamide)

 K+ analysis= Valinomycin

 Hemoglobin A1C (<7%)= DM; monitor Tx

ASCP Pointers- chemistry
 Heparin = green
 Function of wavelength selector

 Instrumentation = photometric, aas,

flurometry
 Phenytoin method measurement

 Volumetric pipet

 Fragile X syndrome

 Bile is tested to detect?

ASCP Pointers- chemistry
 >126 FBS (2x) = DM
 NAD+= urea coupling reactant

 immunoassay test= Creatine kinase (CK)

 p-nitrophenylphosphate = ALP (hepatobiliary)

 Oxidoreductase (oxidase) = uric acid, glucose,

cholesterol and triglycerides
 Low pH, low pO2, increase pCO2 = delay in
blood gas analysis
 Inc CO2, Dec HCO3 = COPD

 Respiratory acidosis
ASCP Pointers
 ROME = respiratory Opposite, Metabolic equal
ph: 7.35-7.45
pCO2 :35-45
HCO3 or ctCO2: 22-26
 Hyperkalemia= metabolic acidosis

 Hyperventilation= respiratory alkalosis

 pH= increased
 pCO2= decreased

 HCO3= normal

Answer: uncompensated respiratory alkalosis

 pH= increased

 pCO2= decreased

 HCO3= decreased

Answer: compensated respiratory alkalosis

pH= 7.52
pCO2 = 20mmHg
HCO3= 24mmol/L
pO2= 100mmHg
The result most likely:
A. Hyperventilation
B. Hyperkalemia
C. Improper specimen collection
D. Improper storage
ASCP question
A patient has a low serum thyroid stimulating
hormone (TSH) and an increased serum T3.
Which of the following result would also be
expected?
A. Elevated free T4
B. Low free T3
C. Low total T4
D. Normal free thyroxine index
When serum potassium and phosphorus are
elevated which one is also expected to rise?
A. Calcium

B. Magnesium
C. AST
D. LD
ASCP Pointers
 PAS (+) = FAB M6 and ALL
 MPO (+), SBB (+), NASDA (+) = FAB M1toM4
 LAP score 10 = CML
 High DIC = FAB M3
 RBC, WBC too blue= poor washing,little buffer
 Target cell, sickle cell, tear drop= thalassemia
 N:C ratio decreases= blood cell maturation
 Decrease nucleoli = blood cell maturation
 Dohle bodies, giant platelets= May-Hegglin
 azurophili granules, mucopolysaccharide defect=
Alder- Reilly anomaly
ASCP Pointers
 Microcytic, hypochromic- iron deficiency
anemia, sideroblastic anemia, thalassemia
 Macrocytic, normochromic= megalobastic
anemia, folic acid deficiency, pernicious anemia
 Normocytic, normochromic= aplastic anemia,
anemia of chronic disease, lead poisoning,
sickle cell disease (hemoglobinopathies) Hgb C,
SC
 Alpha thalassemias= Hgb H & Hgb Barts
 Corrected WBC count = 100 x uncorrected
WBC/ 100 + #nRBCs per 100 WBC
ASCP Pointers
 Rule of 3 = RBC x 3 = Hgb; RBC x 9 = Hct, Hgb
x 3 = Hct
 Hgb F = resist acid elution ( Kleihauer-Betke)

= fetal cells in maternal circulation

 Hgb S = valine replaces glutamic acid

 Hgb C, Hgb E = lysine replaces glutamic acid

 Hgb D = glycine replaces glutamic acid

ASCP Pointers
 No Hgb A = Hgb SS disease, Hgb C, Hgb
SC, Major beta thalassemia
 Hgb electrophoresis = At pH 8.6, Hgb A
migrates fastest, and Hgb A2, C, E, O the
slowest; Hgb S migrates with Hgb D and Hgb G
 Review pictures blast cells

 Teardrop cell picture= myelofibrosis

 (+) PAS = acute erythroleukemia

 ASCP POINTERS
 APTT = Test for Intrinsic factor (8,9,11,12)
and common ( 1, 2, 5, 10)
= heparin and protamine sulfate
 PT (INR) = Extrinsic &common (1, 2, 5, 10, 7)

= Vit. K and coumadin (warfarin)

 TT = FIBRINOGEN

 Bleeding time prolong, APTT high, Restociten

abnormal = von Willebrand
 Inc APTT, Normal PT, Normal BT = Hemophilia
A, VWF Dse, Factor 8, Factor 9
 vWF dse = Inc APTT, Normal PT , Inc BT

 Factor 8 and 9 = Inc APTT, Normal PT , Inc BT

ASCP POINTERS
 Bernard Solier = lack glycoprotein Ib receptor,
Restociten abn.giant platelets, BT prolong,
normal APTT; REVIEW APTT MIXING
STUDY/LAP score
 Platelet satellitism = use sodium citrate not
EDTA
 D dimer (+), FDP (+) = DIC
 D dimer (-), FDP (+) = primary fibrinogenolysis
 Log document control = corrective actions
 Maintenance = quality of machines/equipment
 Computation = Anion gap, Corrective WBC ct
 LAP score is 4+, 3+, 2+, 1+ = leukemoid

 (+) MPO, (+) SBB =APL/ FAB M3

 (+) PAS = ALL

 (+) TRAP = Hairy cell leukemia

 Spherocytes picture

 D dimer = detect fibrin

 Pseudo Pelger Huet = myeloproliferative dse

 BT CT PT APTT Stypven TT Duckert’s

Vascular  N N N N N N
Disoder

Fibrinogen N      N
def.

Prothrombi N     N N
n def
 BT CT PT APTT Stypven TT Duckert’s

Factor VII N   N N N N
def

Classic N  N  N N N
Hemophilia

vWD   N  N N N
 BT CT PT APTT Stypven TT Ducker
t’s
Parahemo N     N N
philia

Hemophili N  N  N N N
aB

Factor X N     N N
def.
 BT CT PT APTT Stypven TT Duckert’s

Hemophilia N  N  N N N
C

Factor XII N  N  N N N
def.

Factor XIII N N N N N N ABN

def.

DIC       ABN
 CYTOCHEMICAL
STAIN

Alpha napthyl
acetate Periodic Acid
Napthol AS-D
Myeloperoxidase Alpha napthyl
Chloroacetate Schiff (PAS)
Sudan Black B butyrate
Esterase (ALL, M6)
(M1-M5) (M4, M5 )
(M1-M4)
(M6,M7
+ Napthylacetate
Cytochemical stain
1. Myeloperoxidase (MPO) stain
 (+) AML (granulocyte, monocyte, Auer rods)
 (-) ALL
2. Sudan Black B ( phospholipids, proteins)
 (+) AML (granulocyte, monocyte, Auer rods)
 (-) ALL
Cytochemical stain
3. Esterases
 Specific esterase stain (napthol AS-D
chloroacetate esterase stain) = granulocyte
 (+) FAB M1, M2, M3, M4
 (-) FAB M5
 Non specific esterase stain (alpha napthyl
acetate and alpha napthyl butyrate) = monocyte
 (+) FAB M5
 (-) FAB M1, M2, M3, M4
Cytochemical stain
4. Periodic Acid Schiff (PAS)
 (+) ALL

 (+) FAB M6 (erythrolekemia)

5. Leukocyte Alkaline Phosphatase (LAP)

 Low LAP score = CML, PNH

 High LAP score = NLR, PV, CML in blast crisis

Cytochemical stain
6. Tartrate Resistant acid Phosphatase (TRAP)
 (+) hairy cell leukemia

7. Perl’s Prussian Blue stain

(+) Siderocytes with iron inclusions
(siderocytic granules/pappenheimer bodies)
(+) HA, beta thallasemia major, sideroblastic Anemia
(+) Sideroblasts = nRBC with iron granules
Ringed sideroblasts= iron encircle nucleus
(+) MDS (Refractory anemia, RARS, sideroblastic
anemia)
Blood chemistry- leukemia
 ALP = CML diagnosis
 Vitamin B12= CML
 Ca, K, phosphate, uric acid = ALL
What clotting factors are inhibited by
coumadin therapy?
A. II and VII

B. II and XI

C. V and VIII

D. XII and XIII

Bleeding time-15 mins (2-4mins)
Platelet count-200 x 109/L
PT- 13 sec (<14 sec)
APTT- 50 secs (20-40 sec)
Abnormal platelet aggregation in response to ristocetin
What is the most probable diagnosis?
A. Bernard-Soulier syndrome
B. Hemophilia A
C. Glanzmann thrombasthenia
D. Von Williebrand’s disease
Which of the following tests evaluates
fibrinolysis?
A. APTT

B. D dimer test

C. PT

D. Fibrinopeptides A and B
A positive non specific esterase stain indicates
differentiation into which cell type?
A. Monocytic

B. Lymphoid

C. Megakaryocytic

D. Plasmacytoid
What condition can be ruled out when positive
to myeloperoxidase stained peripheral blood
smear?
A. Acute myelogenous leukemia M1

B. Acute myelogenous leukemia M4

C. Acute myelogenous leukemia M5

D. Acute lymphocytic leukemia

Which cytochemical stain is used to detect
acute myelogenous leukemia M7?
A. Terminal deoxynucleotidyl transferase (TdT)

B. Myeloperoxidase (MPO)

C. platelet peroxidase

D. Sudan black b (SBB)

Hodgkin Lymphoma
 EBV associated
 (+) Reed Strenberg
cells = large
multinucleated cells
with large nucleoli
 Mild anemia,
eosinophilia,
monocytosis
 High LAP, ESR
Hairy Cell Leukemia (HCL)
 B cell maliganacy (CD
19, CD20)
 Marked splenomegaly
 Pancytopenia
 Cytoplasm show hair
like projection
 (+) TRAP
Hairy Cell Leukemia (HCL)
Chronic Lymphocytic
Leukemia (CLL)
Which of the following stain/reaction show a
positive result in most patient with acute
lymphocytic leukemia?
A. Sudan black and peroxidase
B. Chloroacetate esterase
C. Non specific esterase
D. Terminal deoxynucleotidyl transferase
The leukemia commonly associated with
pediatric age group is:
A. Acute myeloblastic leukemia
B. Acute lymphoblastic leukemia
C. Chronic lymphoblastic leukemia
D. Chronic myelocytic leukemia
All of the following are characteristic of
myelodysplastic syndrome EXCEPT:
A. Lymphocytosis
B. Monocytosis
C. Ringed sideroblast
D. macrocytosis
What is the most common chromosomal
abnormality found in chronic myelogenous
leukemia?
A. t(8;14)
B. t(9:22)
C. t(1;12)
D. Trisomy 12
 That’s all folks….
Remember C A L I
C- Concentration
A- Association
L- Location
I- Imagination