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Packed Cell Volume

Packed cell volume (PCV), also known as hematocrit, measures the percentage of red blood cells in whole blood. It is determined using either the capillary tube method or Wentrob method. The capillary tube method involves centrifuging blood in a capillary tube and measuring the column of red blood cells. The Wentrob method uses an ordinary centrifuge and Wentrob tube. PCV levels are affected by factors like exercise, altitude, patient positioning, and age. The erythrocyte sedimentation rate (ESR) indirectly measures inflammation by determining the rate at which red blood cells settle in a vertical tube. The Westergren and Wintrobe methods are used, differing in tube size and measurement timing

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Motasem Sirag Mohmmed othman
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83 views

Packed Cell Volume

Packed cell volume (PCV), also known as hematocrit, measures the percentage of red blood cells in whole blood. It is determined using either the capillary tube method or Wentrob method. The capillary tube method involves centrifuging blood in a capillary tube and measuring the column of red blood cells. The Wentrob method uses an ordinary centrifuge and Wentrob tube. PCV levels are affected by factors like exercise, altitude, patient positioning, and age. The erythrocyte sedimentation rate (ESR) indirectly measures inflammation by determining the rate at which red blood cells settle in a vertical tube. The Westergren and Wintrobe methods are used, differing in tube size and measurement timing

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Motasem Sirag Mohmmed othman
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MEDICAL LABORATORY TECHNOLOGY 

Packed Cell Volume 


Packed Cell Volume ( PCV )​ ​or​ ​Haematocrit determination, or Hct​.
Haematocrit :​ Is measures the volume (amount) of red blood cells (RBC)
​compared to other components in blood. The results are reported as a
percentage of whole blood.
There are two methods :
1 – Wentrob method .
2 – Capillary tube method .
Capillary tube method​ :
The whole blood separate into 3 areas, ​Plasma​, a ​buffy coat layer​ (i.e.
White blood cells and Platelets), and ​RBCs​ layer.
Principle​:
The blood (mixed with anticoagulant) is placed in a long capillary tube
and centrifuged in a microhaematocrit centrifuge. The level reached by
the column of erythrocytes is read with microhaematocrit reader​.

Materials and reagents​:


1 - Microhaematocrit centrifuge.
2 - Microhaematocrit reader.
3 – Heparinized Capillary tubes ( 75 mm. long with a 1.5 mm diameter,
containing dried Heparin ).
4 – Sealed material Soft wax or clay (or a Bunsen burner or spirit burner).
5 - Sterile blood lancet.
6 - 70% Ethanol​. 
7 – cotton.
8 – blood sample from finger puncture .
Wentrob Method​ :
Materials and reagents​:
1. Ordinary Centrifuge .
2. Wentrobe tube scale from 0 - 10 ( 11 cm. long with 2.5mm diameter,
blood mix with EDTA ).
3. Blood sample from vein .
4. Syringe , Cotton , 70 % alcohol .

Laboratory test results may vary depending on age, sex, health


history, the method used for the test .
(45)
Normal Values : 
1. Adult females : 36 % - 46 % .
2. Adult males : 41 % - 53 % .
3. Infants, 1-2 years : 32 % - 40 % .
4. Children, 9-11 years : 34 % - 43 % .

PCV increased in :  


1. Hard exercise
2. High altitude

PCV decreased in : 
1- Hematocrit is up to 5.7 % lower in recumbent patients.
2- Men aged 65 to 74 years.

Erythrocyte sedimentation rate 


(ESR) 
ESR is a simple non-specific test that indirectly measures the presence of
inflammation in the body. It reflects the tendency of red blood cells to settle
more rapidly in the face of some disease states, usually because of increases in
plasma fibrinogen, immunoglobulins, and other acute-phase reaction proteins,
Changes in red cell shape or numbers may also affect the ESR.
Principle​ : 

When whole blood is mixed with anticoagulant and allowed to stand in a


narrow vertical tube for a period of time, the RBCs – under the influence of
gravity - settle out from the plasma. The rate at which they settle is measured as
the number of millimeters of clear plasma present at the top of the column after
one hour (mm/hr).

There are two main methods used to measure the ESR : ​the Westergren
method​ and the ​Wintrobe Method​ . Each method produces slightly different
results. Most laboratories use the Westergren method.
Westergren method:
The Westergren method requires collecting ​2 ml of venous blood​ into a
tube containing ​0 .5 ml of sodium citrate​. It should be stored no longer than ​2
hours​ ​at room temperature​ or ​6 hours at 4 °C​. The blood is drawn into a
Westergren- tube( ​300 mm long​ , ​2.5 mm. in width​) mark from ​0​ to ​200 mm​.
The tube is placed in a Westergren rack in a strictly vertical position for ​1 hour
at room temperature .
(46)
Wintrobe method : 
The Wintrobe method is performed similarly except that the
Wintrobe tube is smaller in diameter than the Westergren tube and only
100 mm long​. ​EDTA anticoagulated​ blood without extra diluent is
drawn into the tube, and the rate of fall of red blood cells is measured in
millimeters after ​1 hour​. The shorter column makes this method less
sensitive than the Westergren method because the maximal possible
abnormal value is lower.
Some interferences which increase ESR: 

1. Increased level of fibrinogen, gamma globulins.


2. Technical factors : tilted ESR tube, high room temperature.

Some interferences which decrease ESR: 

1- Abnormally shaped RBC (sickle cells, spherocytosis).


2- Technical factors: short ESR tubes, low room temperature, delay in test
performance (>2 hours), clotted blood sample, excess anticoagulant,
bubbles in tube.
There are several factors affect the results of this 
test: 
1. Plasma factors :​ Abnormal increase in some plasma proteins may prevent
the RBCs from falling, causing a falsely decreased ESR. Some abnormal
plasma proteins cause the RBCs to clump quickly and fall at an abnormally
increased rate, resulting in a falsely increased ESR.
2. Red Blood Cell factors :​ Anemia causes a falsely increased ESR​ ​because
the change in the RBC to plasma ration favors rouleaux formation,
causing the RBCs to fall quickly. Rouleaux is condition in which the
RBCs clump together like stacks of coins. Microcytes (abnormally small
RBCs) will fall more slowly than macrocytes (abnormally large RBCs).
RBCs with an abnormal or irregular shape, such as sickle cells (sickle
shaped RBCs) or spherocytes (round RBCs, do not have biconcave
shape), hinder rouleaux formation and cause a falsely decreased ESR.
3. Mechanical and Technical factors :​ Proper technique and correct use of
the equipment is critical for suitable results.

The sedimentation rate of RBCs takes place in 


three stages: 
First Stage -​ ​ Rouleaux formation, the sedimentation rate is slight (20 min.).
Second Stage -​ ​ sedimentation occurs at a fairly rapid rate (20 min.) .
Third Stage ​-​ sedimentation rate is slow because of the accumulation of
RBCs in the bottom of the tube ( 20 min. ).

(47)
Sources of Error: 
1. If the concentration of the anticoagulant is greater than recommended, the
ESR will be erroneously high. EDTA samples may be used for the ESR and
are diluted past to performing the test.
2. If the ESR stands for more than 60 minutes, the results will be falsely
elevated. If the test is timed for less than 60 minutes, falsely low values are
obtained.
3. An increase or decrease in room temperature may lead to increased or
decreased ESR results. The temperature should be within the range of 20-25° C
4.​ The tubes must be maintained in an upright position during the test. All
sedimentation racks used should be equipped with leveling screws and
spirit bubble. Tilting of the ESR tube increases the sedimentation rate.
5.​ Bubbles in the blood tube when filling will lead to erroneous results.
6.​ Fibrin clots present in the blood invalidate the test results.
7.​ The ESR should be set up within two hours of blood collection. EDTA
specimens stored at 4 C for 12 hours or less may be used. The specimen must
be allowed to reach room temperature before the test is performed.
ESR Normal Value : 
Men 0 - 15 mm. per hour (mm. / hr.)
Women 0 - 20 mm. / hr.
Children 0 - 10 mm. / hr.
Newborns 0 - 2 mm. / hr.
 
High ESR may be caused by​ :
1. Autoimmune diseases, such as systemic lupus erythematosus or
rheumatoid arthritis​.
2. Cancer, such as ​lymphoma​ or m ​ ultiple myeloma​.
3. Chronic kidney disease.
4. Infection, such as ​pneumonia​, pelvic inflammatory disease, or appendicitis.
5. Inflammation of joints , and blood vessels .
6. Inflammation of the ​thyroid​ gland (Graves' disease).
7. Kidney​, bone, joint, skin, or heart valve infections.
8. Pregnancy and preeclampsia (toxemia of pregnancy).
9. Viral infections.

Low ESR may be caused by: 


1. High blood sugar levels. 2- ​ ​Polycythemia. 3- ​ ​Sickle cell disease.
2. Severe liver disease.
(48)

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