ERYTHROCYTE SEDIMENTATION RATE - Tubes contain sodium citrate

- Rate at which erythrocytes settle out of 3.2 Sedimat ESR

coagulated blood in 1 hour - Uses the principle of infrared
- NOT SPECIFIC but can detect and measurement
monitor the course of inflammatory - Capable of testing 1-8 samples
conditions randomly or simultaneously and
o Rheumatoid arthritis provides results in 15 minutes
o Infections
o Malignancies 3.3 ESR Stat Plus
- Smaller requires sample volume and
- Diagnosis of temporal arteritis, shorter testing time
polymyalgia rheumatica and rheumatoid - More suitable for pediatric patients
arthritis - Pre-analytical steps must be followed
- Elevated in strictly to prevent erroneous results
o Plasma cell myeloma
o Pregnancy
o Anemia Tube Bore Anticoagu
o Old age length lant
- Prone to technical errors = falsely
elevated or decreased sedimentation Westergren 200 2.5 EDTA or
rate because of low specificity and mm mm sodium
sensitivity citrate 1:4
(black
PRINCIPLE top)
- ESR is the distance in millimeters that Wintrobe 100mm 3mm EDTA or
the RBC fall in 1 hour citrated
- ESR is DIRECTLY proportional to whole
plasma viscosity blood
- RBCs have negative surface charge and
tend to repel one another
o (--)(--) = no attraction; no STAGES OF ESR
rouleux 1. Agglomeration (aggregation)
o (+)(--) = elevated ESR; fast - Initial period about 10 minutes during
attraction which rouleux formation takes place
2. Stage of fast setting (Sedimentation
METHODS rate)
- Period approximately 40 minutes to 1 ½
hour depending on the length of the tube
1. Modified Westergren ESR at which sensitivity or sedimentation
- Most commonly used method occurs at a more less constant state
- Detection of highly elevated ESR 3. Final phase (stationary phase)
- Advantage : taller column height allows - A slower rate of fall during which
the detection of highly elevated ESRs packing of the sediment cells column
occur
2. Wintrobe ESR - 10 minutes
- more sensitive
- shorter column shows increased FACTORS AFFECTING ESR
sensitivity in detecting mildly elevated - RBC
ESRs - Plasma proteins
3. Automated ESR - Mechanical factors
- Depends upon the difference in specific
3.1 Ves-Matic System gravity between the red cells and
- Determine ESR by use of optoelectric plasma
sensor which measures the change in - Actual rate of fall is influenced greatly by
opacity of a column of blood as the extent to which the red cells from
sedimentation of blood progresses

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 rouleaux or aggregate, which sediment
more rapidly than single cells
INTRINSIC FACTORS
1. Number of RBC/mm3
- inversely proportional
- Polycythemia = decreased ESR
- Anemia = increased ESR
2. Size of RBC
- Directly proportional
- Macrocyte = increase ESR
- Microcyte = decrease ESR
3. Viscosity of plasma
- High viscosity = decrease ESR
- Low viscosity = increase ESR
4. Plasma protein content
- Alpha, beta, gamma globulin
- Fibrinogen
EXTRINSIC FACTORS
1. Length of the tube
- Longer tube = increase ESR
2. Diameter
- Wider = increase ESR
3. Position of the tube
- Slightly inclined = increase ESR
4. Temperature (room temperature 25C)
- Increase above 27C
- High temperature = increased ESR
5. Bubbles
- Decreased ESR because of spaces
6. Anticoagulant
- Excess = decrease ESR
7. Time after setting test
- After 2 hours from blood collection =
decrease ESR
8. Time reading results
- Before 1 hour = decrease ESR
REFERENCE VALUES
Males: 0-10 mm/hr
Females: 0-20 mm/hr
PHYSIOLOGICAL VARIATIONS
- ESR is more constant in men than in
women
- In pregnancy, ESR beings to increase at
the 3rd to 4th month and does not retue
to normal until the 3rd or 4th week
postpartum
- In newborn, ESR is greatly reduced; in
older adults it the rather high
COMMENTS AND SOURCES OF ERROR
- ESR is a non-specific indicator of tissue
damage
- Condition in which rouleaux formation is
inhibited, such as sickle cell anemia;
spherocytosis may be accompanied by
a normal ESR or low ESR
- Blood must be fresh. Leaving the
specimen for more than 2 hours at room
temperature will cause RBC to become
spherical and thus inhibiting rouleaux
formation
- Whole blood specimens which are left
overnight will result in lower values
- Slight tilting of sedimentation tube will
increase results
- Presence of anemia invalidates the ESR
as a tool to diagnose a disease process
ZETA SEDIMENTATION RATIO
- Performed using a zeta fuge and special
capillary tubes
- Faster result and not affected by anemia
- Results in 4 minutes
- Specimen: EDTA
- Length: 75 mm long
- Diameter: 2.3 mm (outer) 2.0 mm (inner)
- NV: 40-51%
OSMOTIC FRAGILITY TEST
- Lack sensitivity and non-specific
- Time consuming
- Demonstrates increased RBC fragility in
blood specimens in which the RBCs
have decreased surface-area-to-volume
ratios
- Use graduated concentrations of saline
solutions to detect spherocytes in
hereditary spherocytosis or warm
autoimmune hemolytic anemia
o 0.9 NaCl (isotonic) = normal
RBC
o 1.8 NaCl (hypertonic) = created
RBC
o 0.4 NaCl (hypotonic) = swelling
RBC
- Hereditary spherocytosis – defective
RBC membrane
Increased Decreased
Hereditary Splenectomy
spherocytosis***
Hemolytic anemia Liver disease
Sickle cell anemia
IDA
Thalasemmia
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 METHODS
1. Griffin-Sanford Method
- Uses 12 Wasserman tubes with
decreasing osmolarity of NaCl ranked
from 25 down to 14
- Cells settle at the bottom of the tube and
hemolysis will be recognized by the
color for the supernatant fluid
o Hemolysis:
 Pink – partial
 Red – complete
 Usually begins at test
tube 17
- Specimen: EDTA
2. Rapid screening test
- More rapid ad short-cut modified method
of Griffin-Sanford method
3. Dacie’s Method
- Blood is diluted with isotonic saline
solutions and buffered to ph 7.4 with
phosphate
- Series of tubes is prepared each
containing a different concentration of
saline solution
- Stand at room temperature
- Hemolyzed cells are removed by
centrifugation and the degree of
hemolysis in each tube is determined
spectrophotometrically on the basis of
free hemoglobin
4. Quantitative Method
- Definitive diagnosis of hereditary
spherocytosis
- Normal – slightly increased
- Spherocytosis – greatly increased
5. Unopette Method
- Addition of blood into prepared vials
containing varying concentration of NaCl
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 Points of Differences Wintrobe – Landsberg Westergren
Tube:
Bore (Diamter) 3 mm 2.5 mm
Calibration Up to 100 mm Up to 200 mm
Length 115 mm 300 mm
Anticoagulant Double oxalate (1 part citrate : 3.8% trisodium citrate
4 parts blood)
No. of reading One reading (after 1 hour) Two readings ( after one hour
and two hours)
Normal Values Men: 0-7 mm/hr Men: 2-5 mm/hr
Women: 0-17 mm/hr 7-15 mm/2hr
Children: 1-15 mm/hr Women: 4-7 mm/hr
12-17 mm/2hr
Dilution No dilution Involves dilution (0.6 ml of
3.8% sodium citrate to 1.4 ml
of blood)
Sensitivity Less sensitive More sensitive
Correction of Anemia Applicable Not applicable
Additional tests which may be Hematocrit, micro bilirubin None
performed determination, icterus index
Disadvantages -Not very sensitive because of -Dilution may become source
short column of blood in the of error
tube and the type of -Sensitive but cumbersome
anticoagulant used where -Large amount of blood is
excess oxalate used
-retards ESR -provides no additional
information other than ESR
result

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