Laboratory Evaluation of Platelets

Tuesday, April 10, 2018 7:39 PM

PLT FUNCTION TEST
• Number = Plt Count • Qualitative Plt Disorders
• Function = Adhesion and Aggregations • Normal Plt Count + Bleeding Symptoms = Plt Function Test

PLT COUNT PLT ADHESION

• 150k - 450k /uL • Requires von Willebrand Factor through interaction with glycoprotein 1B
• Thrombocytopenia
○ <150k /uL LABORATORY FUCKING METHOD
• Thrombocytosis 1. Bleeding Time
○ >450k /uL a. Principle: BT is the time it takes for a standard wound to stop bleeding
b. Measures the time it takes for small vessels to close off and stop bleeding
• Plt number must be SUFFICIENT for them to play their supportive c. NV = 2-4 minutes
role in hemostasis. d. Not reliable
i. Use of non-standardized lancet
SUFFICIENT PLT NUMBER ---> EFFECTIVE HEMOSTASIS ii. Pressure

• Bleeding problem = Starts with plt count (starting point) e. Sensitive to abnormalities of : (PVV = Pinoy Vig Vroder)
• There should be 7-20 plts / OIF i. Plt numbers and functions
ii. Vessel Wall
FALSELY LOW PLT COUNT (EGI * EG) iii. vWF deficiencies
• EDTA induced plt satellitism
• Giant Platelets f. Affected by: (QTPM = Cute PM ^^)
• Incipient Clotting i. Quality of Blood Vessels
ii. Thickness of Skin
SPURIOUS ↑ PLT COUNT (Pray MedTech) iii. Plt Count & Plt Function
• Precipitated eryoglobulin (Marked ↑) iv. Medication (Aspirin--> Prolongs BT)
• RBC/WBC Cytoplasmic Fragments
• Microcytic RBC g. Methods: (DITG = Dick In Touch Geez)
• Turbid Plasma i. Duke Method
1) Puncture earlobe, 3rd or 4th finger
SIGNIFICANT PLT COUNT VALUES 2) NV = 1-5 minutes of 2 minutes
3) Wipe the 1st blood with filter paper
<100K / uL Abnormally Low 4) Wipe it every 30 seconds
30k - 80k / uL Bleeding Possible with Trauma ii. Ivy Method
1) It uses blood pressure cuff (sphygmomanometer) inflated at 40
<30K / uL Spontaneous Bleeding Possible
mmHg
<5K / uL Severe Spontaneous Bleeding iii. Template Bleeding Time
1) Modified Ivy Method
PLT ESTIMATE 2) Uses standardized lancet
• Use of blood smear a) 1 mm depth x 5 mm width
• 3-10 plts / 100 RBCs (Note: +2 x2 x2)
• 5-20 plts / 200 RBCs Interpretation:
1) Prolonged in (VVTA= ViVi TAyo)
SIGNIFICANT PLT ESTIMATES a) vWF Deficiency
• Ave # of plts x 20k b) Vascular Disorder
c) Thrombocytopenia
0 49k Marked ↓
d) Acquired and Inherited Plt Disorder
50k 99k Mod. ↓
100k 199k Slightly ↓ Plt Count BT
200k 400k NORMAL Normal Prolonged * Qualitative Disorder
401k 599k Slightly ↑ * Vessel Wall Structure Abnormalities
600k 800k Mod. ↑ Low Normal * Autoimmune thrombocytopenia
>800k Marked ↑ Low Prolonged *Qualitative and Quantitative Disorders

PLT SATELLITISM
• IgG antibody against GP2B3A (Note: GrouP 2 BEA) iv. Glass Bead Retention Time
• Plts surround WBC (Neutrophil) 1) Principle:
• Occurs when EDTA is used a) When blood is passed through a glass bead column, normal
• FALSE ↓ of plt count platelets that have access to normal vWF will adhere and
• To Correct aggregate to the beads such that effluent from the column
○ Use SODIUM CITRATE will have a much lower plt count than the starting sample
○ Multiply plt count by 1.1 b) Second plt count < First plt count
▪ PLT COUNT x 1.1 c) Detained plt = >70% = NV
i) Plt Count 1 - Plt Count 2/ Plt Count 1 x 100
2 TYPES OF PLT COUNT
1. Direct Plt Count PLT AGGREGATION
○ Not Accurate • Requires GP IIb/IIIa
○ Uses RBC pipet, Diluting Fluid or Unopette • Plt Activation Pathway
2. Undirect Plt Count • Function Test: Aids in the Dx of Hereditary and Acquired Plt Disorders
○ Plts are counted in relation to 100 RBCs • Substances causing Plt Aggregation: (EC TARA)
○ Uses Stained Smear ○ Epinephrine
○ Collagen
DIRECT METHOD ○ Thrombin (Human)
1. TOCATIN METHOD ○ ADP
Ristocetin

New Section 1 Page 1

 1. TOCATIN METHOD ○ ADP
a. Reese-Ecker Diluent ○ Ristocetin
i. Makes use of: ○ Arachidonic Acid
1) Sodium Citrate • Dependent on the presence of (FC = Feeling Close)
2) Formaldehyde; Diluent; Fixes and preserves RBC ○ Fibrinogen
ii. Brilliant Cresyl Blue = Stained plts ○ Calcium Ions
• Laboratory Methods
b. Guy and Leake
i. Makes use of: ○ LUMIAGGREGOMETRY / PLT AGGREGOMETRY
1) Sodium Oxalate ▪ Sample:
2) Formaldehyde □ Platelet Rich Plasma from citrated whole blood
3) Crystal Violet  3.8% of trisodium citrate; 1:9 ratio
▪ Centri Speed: 200 x g ; 10 minutes
2. BRECKER CRONKITE ▪ Instrument: Aggregometer
a. Makes use of ▪ Principle:
i. 1% ammonium oxalate (lyses RBC) □ Aggregation agent added to a stired suspension of PRP will induce
ii. Phase Contrast Microscope shape change and aggregation of plts. As a result, the PRP
b. GOLD STANDARD in the direct method changes from turbid suspension to one that transmits more light
c. Plts appear as HALO as the aggregates formed.
▪ Procedure:
3. UNOPETTE METHOD □ Plt Count of PRP: 200-300 x10^9 / L
a. Uses standardized pipette □ Warm 0.5 mL aliquot at 37 Degree Celsius
□ Adjust 100% transmittance of aggregometer
INDIRECT METHOD □ Add the aggregating agent
1. Dameshek
2. Fonio's Method ○ ASPIRIN TOLERANCE TEST
3. Olefs Method - BEST METHOD ▪ 4 tablets = Prolong the BT in NORMAL ptxs
▪ 2 tablets = Prolong the BT in ptxs with vWF deficiencies
AUTOMATED PLT COUNTING ▪ Aspirin - Inhibits Cyclooxygenase enzyme
• Uses electrical impedance/light scatter
• Particles are read as plt if they reach the range of 2-20 fL CLOT RETRACTION
• Forward = Cell Size • Influenced by: (PNC = Pres. Nikko Cataluña)
• Side = Cell granularity ○ P. Cell volume
• MPV = 6-12 fL ○ Number and Activity of Plts
• RBC = 36-360 fL ○ Concentration of Fibrinogen
• Measures serum yield
• Principle:
• Within 1 hour after whole blood is allowed to clot in a clean glass tube @ 37 Degree
Celsius, the clot will begin to shrink and retract

• LABORATORY METHODS (MSC- MS. Cataluña)

○ McFarlane
▪ 5 mL of blood
▪ 37 Degree Celsius for 1 hr
○ Stefanini
▪ 3-5 mL of Blood
▪ Measures at 1 / 2 / 16 / 18 / 24 hrs
○ Castor Oil / Hirschboek
▪ Formation of droplet-like serum on surface
▪ Normal Value = 15-45 minutes

New Section 1 Page 2