HEMATOLOGY 2

LABORATORY // 2ND TERM

BY: MA. ANELY C. BANANG, MD. & MR. JAMES RYAN MENDOZA
1
PLATELET COUNT AND ESTIMATION ○​ Count platelets on a oil immersion objectives
●​ Blood platelets (Thrombocytes) are in 10 fields and multiply by 2,000 which gives
○​ Very small a rough idea of the count
○​ 2-4 um in diameter ○​ Platelets in 10 field x 2,000 = TOTAL PROTEINS
○​ Non-nucleated PLATELET COUNT PRINCIPLE
○​ Membrane bound cell derived from the ●​ Blood is mixed with a diluent in which red cells are
cytoplasm of megakaryocyte of the RBC either left intact or lyzed. A hemocytometer is filled
●​ Each megakaryocyte can produce 2,000-5,000 platelets with diluted fluid and the platelets are counted under
●​ Even platelets like RBC have no nucleus, their the microscope
cytoplasm is packed with granules containing a variety ●​ Whole blood is diluted with Rees-Ecker diluting fluid
of substances that promotes blood clotting which makes platelets readily visible under the
●​ A normal platelet count in human ranges from bright-field microscope
○​ 150,000-400, 000 per microliter of blood ○​ Ammonium oxalate
●​ The life span of platelets is ■​ It can lyse the RBC when testing the
○​ between 8-10 days, platelets we need to see
■​ and those not used in clotting are ○​ Phase contrast microscope
destroyed by macrophages, mainly ■​ Recommended to view using this
in spleen diluting fluid
PLATELET MATURATION ○​ Rees-Ecker
■​ Has 0.1 brilliant cresyl blue that
can't lyse the RBCs which can be
mistaken as platelets
●​ The platelets are often counter in
○​ Central large square of the counting chamber
(hemocytometer)
●​ Collection of blood in the desired volume in EDTA
evacuated tube by venipuncture
○​ Venous blood is preferred
■​ It helps to decrease platelets
clumping, but the MPV (Mean
Platelet Volume) will increase
during the first hour in the tube
PLATELET MORPHOLOGY PRINCIPLE OF DIRECT PLATELET COUNT
●​ Small fragments of cytoplasm derived from ●​ Purpose of performing Total/DIrect platelet count is
megakaryocyte ○​ To know whether the patient is suffering
●​ Stained platelets are lilac from thrombocytosis or thrombocytopenia
●​ Prior to anticoagulation or centrifugation-disk shape (thrombopenia)
with long axis measuring ●​ Specimen is diluted with the diluting fluid which
○​ 2.5-3.5 microns preserves and fix the platelets and stains it
●​ Ability to adhere irreversibly to foreign and ●​ Rees-ecker fluid
endogenously altered surfaces to cohere ( one platelet ○​ Isotonic to Platelets/ Thrombocytes
to another) ○​ Does not cause any damage to whereas
●​ Morphologically three features distinguish by platelet causes the lysis of other cells
from the of other cells ●​ After diluting, the content charged on the
○​ Thick proteoglycans hemocytometer/ neubauer chamber and the cells are
○​ Ability to form fibrillar bridges between one counted in the specific areas for platelets count (The
platelet membrane to another four "WS")
○​ Distribution of intramembranous particles REAGENTS AND MATERIALS
which differs from that of any mammalian ●​ EDTA Evacuated Tube
cell ●​ Microscope: LPO and HPO
METHODS FOR COUNTING PLATELETS ●​ Diluting Fluid: Rees-Ecker Fluid
●​ By automated hematology analyzer ○​ Composition
●​ Direct smear ■​ 3.8 g of Trisodium citrate
○​ also gives information about platelet size, ●​ Prevents coagulation
shape and clumping clumping of Platelets
●​ Direct count from the peripheral blood smear ■​ 0.2 mL Neutral Formaldehyde
●​ Preserve the pits

ALE, D.A. | MED 223

HEMATOLOGY 2
LABORATORY // 2ND TERM
BY: MA. ANELY C. BANANG, MD. & MR. JAMES RYAN MENDOZA
2
■​ 0.1 Brilliant Cresyl Blue
●​ Stains the pits
■​ 100 mL Deionized water
●​ Hemocytometer and coverslip
●​ RBC pipette with rubber tubing and mouthpiece
●​ Tally or Cell Counter
PROCEDURE
●​ The capillary bore of the RBC pipette is rinsed with
Rees-Ecker diluting fluid.
○​ All excess fluid should be removed from the
pipette before blood is drawn into the pipette
before blood is drawn into the pipette.
○​ This step will ensure platelet will not adhere
on the wall of the capillary bore
●​ Draw the blood up to 0.5 mark
○​ If excess blood was drawn, adjust the blood
level to the exact 0.5 mark with NSS-
moistened cotton
○​ Clean the stem of the pipette first before any
adjustment RBC PIPETTE WBC PIPETTE

●​ Blood should be diluted immediately with the

BEAD COLOR RED WHITE
Rees-Ecker solution.
○​ Make a 1:200 dilution by filling the pipette GRADUATIONS UP TO 101 MARK UP TO 11 MARK
with the solution up to 101 mark
○​ Two pipettes should be used to BULB LARGER SMALLER
simultaneously, and each should be used to COUNTING SYSTEM TO ENSURE ACCURACY AND CONSISTENCY
charge one side of the counting chamber
●​ If 5 RBC squares are counted area should be 0.20 mm²
●​ After the dilution, the pipette should be shaken
●​ If 25 RBC squares (central large square) are counted
immediately at least 60 seconds
area should be 1 mm
○​ This will prevent platelet clumping and will
ensure accurate counting
●​ After mixing, discard the first 5 drops from each pipette
○​ Note: Do not induce or blot the tip of the
pipette by any absorbent material to facilitate
the process. The drop should drop freely by
gravity alone. Each side of the
Hemocytometer should be charged with a
different pipette
●​ After charging the chamber the Hemocytometer should
be kept in covered for about 10-15 minutes with wet
gauze or cotton pads beneath the hemocytometer
○​ To prevent evaporation (drying up)
○​ This step will allow the platelets to settle in CALCULATIONS
accurate counting later on ●​ As the RBC and WBC counts, the important parameters
●​ Carefully place the hemocytometer or neubauer on the for calculating the platelet count includes
microscope stage so as not to disturb the platelets. ○​ The average number of platelets per square
○​ Count plates in 5 RBC squares in the center of milimeter
the counting chamber ○​ The dilution factor (which is 200 um like the
○​ Both sides of the chamber should be counted RBC count)
and the results averaged ○​ The volume of the diluted blood counted
●​ Using the high-power magnification to count all the which is the area times depth (1 mm x 0.1
platelets, they appear as small, roundish, unevenly mm = 0.1 uL)
shaped structures

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 HEMATOLOGY 2
LABORATORY // 2ND TERM
BY: MA. ANELY C. BANANG, MD. & MR. JAMES RYAN MENDOZA
3
○​ Platelets adhere to neutrophil when the blood
sample is anticoagulated with EDTA and not
free to be counted
●​ Reference Range: ●​ Corrected by re-drawing blood sample using sodium
○​ 150 - 450 x 10⁹/ L citrate as anticoagulant and the resulting platelet
●​ Note: count should be multiplied by 1.1 to account for
○​ RBC Pipette: 2-3 drops delusional effect of citrate
○​ WBC Pipette: 1-2 drops SOURCE OF ERROR
CHARACTERISTICS OF A GOOD PLATELET DILUTING FLUID ●​ Inadequate mixing and poor collection of the specimen
●​ Should prevent platelets from adhering to glass surface can cause the platelets to clump on the hemocytometer.
of the pipette If the problem persists after resolution, a new
●​ It should also prevent the platelets from aggregating specimen is needed. A skin puncture specimen is less
●​ Should not promote premature platelet hemolysis desirable because of the tendency of the platelets to
●​ Should make platelets readily visible under a aggregate or form clumps
microscope ●​ Dirt in the pipette, hemocytometer, or diluting fluid
REES-ECKER DILUTING FLUID COMPOSITION may cause the counts to be inaccurate
●​ Sodium Citrate ●​ If fewer than 50 platelets are counted on each side, the
○​ Prevents coagulation procedure should be repeated by diluting the blood to
●​ Formalin 1:20
○​ Fixes Platelet and thus prevent premature ●​ If more than 500 platelets are counted on each side, a
hemolysis 1:200 dilution should be made. 1:200 dilution should be
●​ Brilliant Cresyl Blue made. The appropriate dilution factor should be used
○​ Allows the platelets to be observed under a in calculating the results
microscope ●​ If a patient has a normal platelet count
NORMAL PLATELET COUNT ○​ The 5 small, red blood cell squares may be
counted.
AGE cmm (mm³) X10/ L (SI UNIT) ○​ Then, the area is 0.2 mm² on each side
●​ The phenomenon of "Platelet Satellitosis" may occur
Adult/ Elderly 150,000 - 400, 000 150 - 400
when EDTA anticoagulant is used. This refers to the
Premature infants 100,000 - 300,000 100 - 300 adherence of platelets and neutrophils, producing a
ring or satellite effects
Newborns 150,000 - 300,000 150 - 300 ○​ Using Sodium Citrate as the anticoagulant
should correct this problem. Because of the
Infants 200,000 - 475,000 200 - 475 dilution in the citrate evacuated tubes, it is
necessary to multiply the obtained platelet
Children 150,000 - 400, 000 150 - 400
count by 1.1 for accuracy
●​ Thrombocytopenia PERIPHERAL BLOOD SMEAR
○​ Less than 100,000 cmm ●​ EDTA Blood
●​ Thrombocytosis ○​ Anticoagulant of choice in platelet counting
○​ More than 400,000 cmm ○​ Preserves morphology of platelet counting
●​ Thrombocythemia ○​ Distinguishable platelets; no significant
○​ Above 1 million cmm change in shape or appearance
THINGS TO REMEMBER ●​ Glass slide
●​ Platelet count should be performed within 3 hours after ●​ Wright Stain
after the dilution has been prepared PLATELET COUNT
●​ If the blood is collected by a skin puncture, carefully ●​ Microscope
remove the first drop of blood and collect free flowing ●​ Immersion oil
blood for the platelet count. This will minimize
occurrence of platelet clumping and adhesion of
platelets to the puncture site
●​ If clumps are present, procedure should be repeated
●​ Wright stained peripheral blood smear is preferred
○​ Should be examined and the platelet estimate
determined to confirm the hemocytometer
platelet count
●​ Platelets satellitism will result in Falsely Decreased
platelet counts

ALE, D.A. | MED 223

HEMATOLOGY 2
LABORATORY // 2ND TERM
BY: MA. ANELY C. BANANG, MD. & MR. JAMES RYAN MENDOZA
4

ZONE OF MORPHOLOGY
●​ Are where you evaluate morphology of Red Cells
●​ Where we start Differential Count
●​ Where we start Counting Platelets
ESTIMATED PLATELET COUNT
●​ Derived by correlating a machine count and a clear
count
●​ 30 specimens from health subjects
○​ Machine count
○​ Prepare PBS for each
CLINICAL SIGNIFICANCE
●​ Increased platelet count
○​ Polycythemia Vera
○​ Idiopathic Thrombocytopenia
○​ Chronic Myelogenous Leukemia
○​ Splenectomy
●​ Decreased platelet count
○​ Thrombocytopenia Purpura
○​ Aplastic Anemia
○​ Gaucher's Disease
○​ Pernicious Anemia
○​ Chemotherapy
○​ Radiation Therapy

ALE, D.A. | MED 223

HEMATOLOGY 2
LABORATORY // 2ND TERM
BY: MA. ANELY C. BANANG, MD. & MR. JAMES RYAN MENDOZA
5
CAPILLARY FRAGILITY TEST ■​ Male hemophiliacs cannot undergo
●​ Intentionally inducing trauma to test the ability of circumcision
blood vessels to maintain tenacity under pressure ■​ Hemophiliacs can undergo tooth
PROCEDURE extraction and other minor
1.​ Draw a 5 cm ring on a patient's forearm with a 4 cm surgeries but require a
space from the skin fold multidisciplinary approach
2.​ Apply the BP cuff above the skin fold. Pump pressure at PLATELET FUNCTIONS UNDER NORMAL HEMOSTASIS
80mmHg and maintain for 8 minutes
ANTICOAGULATION PROCOAGULATION
3.​ Remove the BP cuff after 8 minutes
4.​ Wait 5 minutes, afterward, count the number of Normal Abnormal
formed petechiae inside the 5cm ring
CONDITION Intact Vessel Integrity Damaged Vessel Integrity
IDEAL
●​ Choose a forearm without existing petechiae or
discoloration
PRIMARY HEMOSTASIS SECONDARY HEMOSTASIS
ACTUAL
●​ Count existing petechiae and if both arms have
Platelet Plug Formation Fibrin Mesh Formation
petechiae – choose the arm with a lesser presence of
petechiae
REPORTING CAUSES OF THROMBOCYTOPENIA

●​ Reference ●​ Viral Infection

○​ <10 petechiae ○​ Dengue fever

■​ Acquired from Aedes aegypti
mosquitoes
Score Number of Formed Petechiae
●​ Idiopathic/ Unknown Causes, Autoimmune Causes
○​ Henoch-Schonlein Purpura
1+ 1 - 10
■​ A condition where blood vessels
2+ 11 - 20 become inflamed
■​ Causes splenic sequestration of
3+ 21 - 50
platelets

4+ 51 - higher HEREDITARY DISEASES THAT CAUSE FRAGILE BLOOD VESSELS

●​ Ehlers-Danlos Syndrome
○​ A group of disorders that affects connective
CLINICAL SIGNIFICANCE OF CFT
tissues
●​ Assessment of vascular integrity
○​ Characterized by flexible joints
●​ Assessment of risks for mucocutaneous bleeding
(hyperextension) & retractable skin
○​ Petechiae
○​ Purpura
○​ Ecchymoses
MOST COMMON PATHOLOGIC CASES OF CFT
●​ Thrombocytopenia
○​ Decreased PCT
●​ Scurvy
○​ Vitamin C Deficiency:
■​ Vitamin C encourages collagen
production in the tissue matrix
■​ ↓ Vitamin C = ↓ Collagen
■​ Pirates were commonly diagnosed
with Scurvy due to the
inaccessibility of Vitamin C
■​ Vitamin C (ascorbic acid) is
prescribed to patients who
underwent c-section to promote
collagen production and faster
recovery
●​ Hemophilia
○​ Decreased Coagulation Proteins

ALE, D.A. | MED 223

HEMATOLOGY 2
LABORATORY // 2ND TERM
BY: MA. ANELY C. BANANG, MD. & MR. JAMES RYAN MENDOZA
6
PROTHROMBIN TIME, INR, SIS FACTOR VII
●​ Report time or clot ●​ Stable factor (3-6 hours)
●​ Report INR REAGENTS AND EQUIPMENT
●​ Report International Sensitivity Index ●​ Thromboplastin
PROTHROMBIN TIME (PT) ○​ Rabbit brain or lung tissue
●​ Use screening procedure for the extrinsic coagulation ●​ Calcium chloride (CaCl₂)
mechanisms including the common pathway ○​ Keep refrigerated when not in use
○​ Detects deficiencies in Factors: ●​ Normal and Abnormal plasma controls
■​ II ●​ Test tubes
■​ V ○​ 13 x 100 mm
■​ VII ●​ Stopwatch/ Timer
■​ X SPECIMEN
●​ Used to monitor oral anticoagulants ●​ Citrated plasma
○​ Warfarin ○​ Testing should be performed within 2 hours
○​ Coumadin after specimen collection when the plasma is
○​ Courmarin maintained at room temperature and within
●​ The prothrombin time (PT) in an individual with one or 2 to 5 days when the plasma is stored at -20°C
more deficiencies of clotting factors will vary with the PRINCIPLE
type of thromboplastin (e.g. rabbit, human, bovine)
●​ The calcium in whole blood is bound by sodium citrate,
used in the assay. This difference in sensitivities is
thus preventing coagulation. Tissue thromboplastin, to
known as the sensitivity index
which calcium has been added, is mixed with the
●​ Normal range:
plasma, and the clotting time is noted
○​ 10-14 seconds
●​ Factors VII reacts with the tissue factor
●​ These values differ according to the method and
(thromboplastin) in the presence of calcium ions to
reagents and lot numbers used in the performance of
activate factor X. The coagulation mechanism
the test and must be determined by each laboratory
continues from there
●​ As a general rule, only 30% concentration of Factor V,
PROCEDURE
VII and are needed for a normal PT
●​ Centrifuge citrated sample to obtain Platelet Poor
PROLONGED PT
Plasma (PPP)
●​ Vitamin K Deficiency (1972)
●​ Testing should proceed immediately or cap the
○​ Bile duct atresia
centrifuged or plasma tube and complete the procedure
○​ Irresponsible antibiotic intake
within 2 hours
○​ Maternal antibodies for breastmilk
●​ Pipette 0.2 mL of thromboplastin-calcium reagent into
●​ Certain liver diseases such as severe liver damage
the appropriate number of 13 x 100 mm test tubes.
○​ Cirrhosis
Warm the test tubes in the water bath for at least 1
●​ Specific coagulation deficiencies
minute until they reach 37°C
○​ Factor VII
○​ The incubation period for this mixture is not
●​ Disseminated Intravascular Coagulation (DIC)
critical once in reaches 37°C
○​ Fibrinolytic protein
●​ Incubate the plasma for 2 to 3 minutes, until it reaches
■​ C
37°C
■​ S
○​ Plasma should be incubated no longer than 5
■​ Z
minutes after reaching 37°C
●​ Circulating anticoagulants
●​ Forcibly pipette 0.1 mL of patients plasma into the test
○​ Lupus anticoagulant targets thromboplastin
tubes containing 0.2 mL of thromboplastin-calcium
●​ Presence of fibrinogen split products
mixture and simultaneously start the stopwatch
●​ Mix the contents of the tube, remove the tube from the
FIBRIN FIBRINOGEN water bath, and wipe dry. Gently tilt the tube back and
forth until a clot form, at which point the timing is
D - DIMER NO D - DIMER stopped
●​ Each test and control plasma should be performed in
●​ Some dysproteinemia duplicate when manual methods are used
○​ 3.2 Na Citrate ○​ The results should agree within +/- 0.5
○​ Forms soluble Ca complexes seconds when the prothrombin time is below
●​ Oral anticoagulant therapy 30 seconds
○​ Warfarin therapy ○​ Duplicate test on prothrombin time above 30
●​ Dysfibrinogenemia seconds should agree within 2-4 seconds,
depending on the degree of prolongation

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HEMATOLOGY 2
LABORATORY // 2ND TERM
BY: MA. ANELY C. BANANG, MD. & MR. JAMES RYAN MENDOZA
7
○​ Average the two clotting times and the report
the patient's results within the normal range
for the test as performed in your laboratory
●​ Standard Centrifugation time
○​ 15 minutes
INTERNATIONAL SENSITIVITY INDEX (ISI)
●​ Individual Thromboplastin can be calibrated against
an International World Health Organization reference
Thromboplastin (International Reference preparation
for IRP) to assign then an International Sensitivity
Index (ISI)
●​ The first WHO references Thromboplastin was
assigned an ISI of 1.0 and it is against this (and
subsequent reference preparations) that all other
Thromboplastin are calibrated
●​ The first WHO IRP was a human brain extract to which
adsorbed bovine plasma was added to optimize the
content of the non-vitamin K dependent coagulation
factors – adsorption with Al(OH)³ removes all the
vitamin K dependent proteins
●​ Subsequent WHO IRPs contain no adsorbed bovine
plasma
INTERNATIONAL NORMALIZED RATIO (INR)
●​ The calculation made to standardized prothrombin
●​ It is based on the ratio of the patient's PT and normal
mean PT
●​ Is the PT ratio of a test sample compared to a normal
PT (derived from the log mean normal prothrombin
time (LMNPT) of 20 normal donors) corrected for the
sensitivity of the Thromboplastin used in the test
●​ It is the value for the Prothrombin Time Ratio which
has been obtained using the first WHO Reference
Thromboplastin with an ISI of 1.0.

ALE, D.A. | MED 223

 HEMATOLOGY 2
LABORATORY // 2ND TERM
BY: MA. ANELY C. BANANG, MD. & MR. JAMES RYAN MENDOZA
8
ACTIVATED PARTIAL THROMBOPLASTIN TIME (APTT) ●​ Hematemesis
USES OF ACTIVATED PARTIAL THROMBOPLASTIN TIME ○​ Vomiting of blood
●​ Evaluation of intrinsic pathway of Coagulation ●​ Menorrhagia
●​ Liver Disease ○​ Profuse menstrual flow
○​ Multi-factor deficiency ●​ Hemarthrosis
●​ Hypofibrinogenemia ○​ Joint bleeding causing acute pain and
●​ Heparin monitoring swelling
●​ Von willebrand disease MIXING STUDIES/ SUBSTITUTION TEST
○​ Variable ●​ Are test performed on blood plasma used to distinguish
REAGENTS AND MATERIALS FOR APTT factor deficiencies from factor inhibitors
●​ Thromboplastin ○​ Lupus anticoagulant
●​ Calcium chloride ○​ Anti-factor VIII
●​ Test tubes ●​ Also referred to as circulating inhibitor screen
●​ Pipette ●​ Patient's plasma is diluted with normal pooled plasma
●​ Water bath or dry bath demonstrated factor levels
●​ Timer ●​ The normal plasma provides the missing factor in the
●​ Specimen patient plasma
○​ Citrated poor plasma
PROCEDURE
Extrinsic Intrinsic Normal Adsorbed Aged Serum
●​ Pre-warm a sufficient quantity of calcium chloride Pathway Pathway Plasma Plasma
reagent to 37°C for at least 10 minutes
I I + + -
●​ Pipette 100 uL of sample into a labeled test tube,
incubate 1 to 2 minutes
II II + - -
●​ Into each test tubes, add 100 uL of partial
thromboplastin reagent V V + + -
●​ Incubate the mixture at 37°C for a minimum of 3
VII + - +
minutes
○​ Optimum activation of contact factors
VIII + + -
●​ Reference value
○​ 25 to 35 seconds XI + - +
FACTOR DEFICIENCY EVALUATION
x X + - +
●​ First consideration
○​ PT and/or APTT must be prolonged XI + + +
○​ Patient history and symptoms must be
considred XII + + +

●​ Follow-up tests ADSORBED PLASMA

○​ Mixing studies PROCEDURE
BLEEDING DISORDER MANIFESTATION 1.​ Blood samples are taken from 10 healthy donors in
Primary Hemostasis Disorder Secondary Hemostasis Disorder trisodium citrate tubes
(Mucocutaneous Bleeding) (Anatomic/ Soft Tissue Bleeding) 2.​ Platelet poor plasma is made by centrifuging each
sample at 400 rpm/min for 15 minutes
Petechiae Excessive/ recurrent bleeding
3.​ Prothrombin time is noted
Purpura after minor trauma, dental
extraction and surgery a.​ Normal samples are separated
4.​ Samples with normal values are poured in a test tube,
Ecchymosis Hemarthrosis thus making a pool
Gum bleeding Bleeding involving body cavities
a.​ Volume is noted in mL
Hematemesis
Menorrhagia 5.​ Barium sulphate is weighed at 100 mg/mL pool plasma
6.​ Barium sulphate is mixed frequently in the pool
●​ Petechiae
plasma by glass rod in water bath at 37°C
○​ Pinpoint hemorrhages into the skin less than
7.​ After 15 minutes, a slow spin at 1000 rpm/min is given
3 cm
to the pool plasma for 3 minutes
●​ Purpura
a.​ Barium sulphate settles down
○​ Purple lesion of the skin greater than 3 mm
8.​ Prothrombin time is performed on the supernatant
●​ Ecchymoses/ Bruises
9.​ If it is 60 to 80 seconds, the process is complete:
○​ Greater than 1 cm seen after trauma
a.​ Store the adsorbed plasma at -30°C in labelled
●​ Epistaxis
cuvette
○​ Uncontrolled nosebleed
10.​ If it is <60 seconds

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 HEMATOLOGY 2
LABORATORY // 2ND TERM
BY: MA. ANELY C. BANANG, MD. & MR. JAMES RYAN MENDOZA
9
a.​ Add more Barium sulphate
i.​ E.g. 10 mg and start from step 6
11.​ If it is >80 seconds
a.​ Add more pool plasma
i.​ Already separated aliquots and also
start from step 6
12.​ Ensuring writing prothrombin time and date of
manufacturing on the cuvette or the container having
cuvettes
PROCEDURE OF MIXING STUDIES
1.​ Prepare 1:1 ratio of patient's samples and normal
plasma/ adsorbed plasma/ aged serum
2.​ Determine PT and APTT level
3.​ If corrected
a.​ Factor deficient
4.​ If uncorrected
a.​ Presence if circulating inhibitor
CIRCULATING INHIBITOR SCREEN
●​ Lupus anticoagulant/ Antiphospholipid
○​ Non-specific inhibitor, have different targets
○​ Should be suspected in a patient with marked
prolonged PT and APTT but no clinical
symptoms
○​ The abnormal antibody reacts mildly in vivo
with thrombotic events, but reacts stronger
in vitro by bleeding to the phospholipids in
the reagents used for coagulation testing
○​ Directed to phospholipid
●​ Anti-factor VIII
○​ Most common specific inhibitor for
hemophiliacs (10 to 20%)
●​ Anti-factor IX
○​ 1 to 3% in factor IX deficient patients

DEFICIENCY OR IMMEDIATE PR OR MIXING STUDY

INHIBITOR APTT AFTER MIXING FOLLOWING 2 HOUR
STUDY INCUBATION

Factor Deficiency correction correction

Lupus-like no correction no correction

anticoagulant

Factor VIII inhibitor correction no correction

ALE, D.A. | MED 223