What is one of the most complex areas of diagnostic laboratory testing?
Hemostasis
What are the 3 steps for forming a clot?
1. constrict the blood vessel in the region of the hole
2. form a platelet plug
3. seal that plug with fibrin
What is platelet poor plasma?
contains less than 10 x 109 platelets/L
What are some specimen factors affecting coagulation testing?
- collection system
- order of draw
- anticoagulant
- phlebotomy technician
- centrifugation
- delays in handling
- storage conditions
- order of draw
What are some instrument factors affecting coagulation testing?
- temperature
- light sensor
- detector
- timer
- disposables
- reagent delivery
What are some reagent and control factors affecting coagulation testing?
- shipping conditions
- storage conditions
- reconstitution
- contamination
- deterioration
- lot changes
What must be analyzed with each assay performed within the lab?
Internal Quality Control (IQC)
What is the purpose of internal quality control?
To assess ongoing assay performance to ensure assays are:
- performing according to specifications
- results are accurate and reliable
- ensure reproducibility of testing
Definition:
Peer group assessment process that allows a lab to assess individual analytes against those of other labs using either the same or different reagents or instrumentation
External Quality Assurance
What anticoagulant is used for routine coagulation testing?
Sodium citrate
True or False:
If sodium citrate anticoagulant is unavailable, you may use EDTA, heparin, or oxalate.
False, these anticoagulants are unacceptable
What is the ratio of blood to anticoagulant?
9:1
A disproportion of blood to anticoagulant is seen in a patient with what disorder?
Polycythemia
What is the calculation used to calculate the amount of sodium citrate that should be used?
- X= (100-Hct)Vol/(595-Hct)
- X = volume of sodium citrate required for unit volume of blood
- Hct = hematocrit
- Vol = blood draw volume required in the blood collection tube
Most clot based coagulation activity procedures are performed on what type of specimen?
Plasma
The addition of what to plasma is required in order to perform coagulation activity testing?
The addition of CALCIUM is required in order to perform tests
What pathway include factors X, V, II, and I?
Common pathway
Which pathway includes factor VII?
Extrinsic pathway
Which pathway include factors XII, XI, IX, and VIIIa?
Intrinsic pathway
What are some advantages of enzyme specific synthetic substrates over the traditional clot formation?
- can be performed rapidly
- sensitive
- allow for a greater degree of standarization
- require smaller sample volumes
- well suited for automation
If unable to perform testing within 4 hours what should be done?
Freeze plasma at -70C
Immediately before testing plasma, how should you thaw it when frozen?
Rapidly thaw in a 37C water bath to prevent formation of cryoprecipitate
What are the 4 methods of endpoint detection?
- Mechanical
- Photo-optical
- Chromogenic
- Immunogenic
A fibrometer is what type of endpoint detection method?
Mechanical
What type of endpoint method is the following:
when a light souce of a specified wavelength is passed through a test solution (plasma), a certain amount of light is detected by a photodetector or photocell located on the other side of the solution
Photo-optical endpoint detection
This endpoint detection method is the most commonly utilized in coagulation instruments
Photo-optical endpoint detection
What is another name for photo-optical endpoint detection?
Turbidometric methodology
This endpoint detection method is also known as amidolytic methodology
Chromogenic Endpoint Detection
The color-producing substance used in chromogenic endpoint detection is known as what?
Chromophore
This endpoint detection method is based on antigen-antibody reactions
Immunogenic endpoint detection
What bings von Willebrand Factor (vWF)?
Glycoprotein Ib
What binds fibrinogen?
Glycoprotein IIb-IIIa
These granules inside platelets are also known as specific granules and contain fibrinogen, von Willebrand factor, platelet-derived growth factor, and P-selectin
alpha granules
These granules are known as dense granules and contain serotonin, ATP, ADP, and calcium (which is necessary for the vitamin K dependent coagulation factors)
delta granules
What type of proteins help the platelet change shape during adhesion?
Contractile proteins (like actin)
What are the 3 steps of platelet plug formation?
1. adhesion
2. activation
3. aggregation
Definition:
measured as a screening procedure used to detect both congenital and acquired disorders of platelet function and von Willebrand disease..
Bleeding Time
Whare are the 4 procedures in use for determining bleeding time?
- Duke method
- Ivy method
- Mielke method
- Surgicutte or Simplate method
This bleeding time method punctures the ear lobe
Duke method
This bleeding time method requires a blood pressure cuff to maintain constant pressure within the capillaries to help standardize the procedure. The forearm is the site used and a lancet is used to make the incision. The greatest source of variation is in performing a standardized puncture.
Ivy method
This bleeding time method is a modification of the Ivy method. A disposable blade (scalpel) is used.
Mielke method
This bleeding time method is the preferred method. The device used has a trigger and spring method for the blade to make a precise 1.0 mm depth cut with a width of 5.0 mm.
Surgicutte/Simplate method
During the bleeding time procedure, the filter paper is slid into the droplet of blood how often until bleeding ceases?
Every 30 seconds
During the bleeding time procedure, the sphygmomanometer is kept at what measurement?
40 mmHg
What is the normal range for bleeding time?
2.5-9.5 minutes
What may cause a porlonged bleeding time?
- The use of aspirin, aspirin-containing drugs, non-steroidal anti-inflammatory drugs, and antihistamines
- If the alcohol used to clean the puncture site isn't fully dried
- If the technician allows the filter paper to touch the wound, the platelet may dislodge
Obtaining an accurate bleeding time may not be possible in what type of patients?
Older patients because bleeding may only occut subcutaneously
True or False:
Thrombocytopenia with a platelet count less than 50,000/uL may cause a shortened bleeding time
False- it would cause a prolonged bleeding time because there are less platelets to form the plug
What are some inherited platelet dysfunctions that would cause prolonged bleeding times?
Bernard Soulier syndrome
or
Glanzmann's thromboasthenia
or
Storage pool disease
True or False:
Von Willebrand disease causes a shortened bleeding time
False
Von Willebrand disease causes a prolonged bleeding time