Esr and its methods

1. Arun Kumar.
II M.Sc Biochemistry.

2.  It is a group of inherited red blood cell disorder.(Autosomal
Recessive)
 Healthy red blood cells are round and they move through
small blood vessels carrying oxygen to all parts of the body.
 In sickle cell disease, the red blood cells become hard and
sticky and look like a C-shaped farm tool called a sickle.
 Sickle cells die early, which causes a constant shortage of red
blood cells.
 The Glutamic acid in the 6th position of beta chain of HbA os
changed to Valine in HbS.
 This causes distortion of cell into sickle shape.
 The substitution of hydrophilic glutamic acid by hydrophobic
valine causes localized stickness on the surface of the
molecule.

4.  Sickle cells can get stuck in small blood vessels and block the
flow of blood and oxygen to organs in the body. These
blockages cause repeated episodes of severe pain, organ
damage, serious infections, even stroke.

6.  Sickle cell anemia:
Homozygous state for HbS(βS-βS)
 Sickle cell trait:
Heterozygous carrier state for HbS(βS-β)
 If one parent has sickle cell anemia and other is normal, all
children will have sickle cell trait.
 If one parent has sickle cell anemia and other has sickle cell
trait there is 50% chance of either with each pregnancy.

9.  New Born spot screen.
 Blood smear
◦ Sickled cells.
 Protein Electrophoresis.
 Erythrocyte Sedimentation Rate [↓ed]
◦ Non-Specific Test.
(because haemoglobin is in sickle shape so it
agglutinates and sediments slowly)

11.  Erythrocyte sedimentation rate (ESR) is a nonspecific test for
inflammation.
 It is easy to perform, widely available and inexpensive making
it a widely used screening test.
 It is also used a monitoring tool for response to treatment in
conditions in which it is raised (Tuberculosis, Sickle Cell
Anemia, Autoimmune diseases etc..)
 Consist of allowing a specific amount of blood to sit in a
vertical position for a period of time (usually one hour).
 The distance, in millimeters, that the red cells fall during this
time period is the erythrocyte sedimentation rate and is
reported in mm at one hour.

12.  Rouleaux formation (10 minutes) Red cells
stack together like pair of coins
 Sinking of rouleaux (40 minutes)
 Cell packing : (10 minutes)

13. 1. Erythrocytes:
 Macrocytes tend to sediment rapidly than
microcytes.
 Alterations of ratio of red cells to plasma
 Decreased red cell mass in anaemia increases ESR .
 Increased red cell mass in polycythemia & Sickle Cell
Anemia decreases ESR.
 Sickle cells and spherocytes are unable to form
rouleaux and therefore ESR is low.
2. Plasma Composition:
 In normal blood, the RBCs remain more or less separated.
They are negatively charged and, therefore repel each
other. ( Zeta Potential )

14.  Increased plasma protein concentration, most notable
fibrinogen, immunoglobulins and acute phase proteins (
Creactive protein, ceruloplasmin, -1 globulin) , cause a
reduction in the negative charge of the RBCs and facilitates
formation of rouleaux.
 Removal of fibrinogen by defibrination and increase in the
albumin retard ESR.
3. Mechanical/Technical factors:
It is important that the ESR tube be exactly perpendicular.
A tilt of 30⁰ can cause errors up to 30%.
The rack, holding the tubes should not be subject to any
movement or vibration.
The sedimentation rate increases as the temperature
increases.
ESR tubes with a narrower than standard bore will
generally yield lower sedimentation rates.

15.  MANUAL –
Wintrobe’s Method .
Westergen’s Method.
Zeta Sedimentation ratio.
Micro ESR Method.
 AUTOMATED –
Monitor 100
Ves-matic 60
Sedimatic
Sediscan

18.  Mix the anti coagulated blood thoroughly.
 Fill the wintrobe tube by using Pasteur pipette
upto mark 0.
 Place the tube vertically in a stand.
 Note the ESR at the end of 1 Hour.

21.  Venous blood collected in trisodiumcitrate
solution in 4:1 proportion
 Test should be carried out within four hours
of blood collection if specimen is kept at
room temperature
 Mix the blood and anticoagulant thouroughly.
 There should be no clots and air bubbles in
the blood.

22.  Fill the Westergren pipette by inserting in a
vacutainer tube containing 1.6 ml blood and
0.4ml anticoagulant
 Keep the pipette upright in the ESR stand
lying on the leveled surface
 Read the upper level of RBC column exactly
after one hour.

23.  The ESR represents a nonspecific response to tissue damage
and inflammation.
 It primarily reflects changes in the plasma proteins that
accompany most the acute and chronic infections, tumors,
and degenerative diseases.
 It may be used to follow the progress of certain diseases such
as tuberculosis and rheumatoid arthritis.
 Important criterion in establishing the diagnosis of temporal
arteritis and polymyalgia rheumatica.

24. INCREASE IN ESR DECREASE IN ESR
-Most Anemia. -Sickle cell anemia.
-Inflammation or infection. -polycythemia.
-Cancers, ESRD, or pregnancy. -Heart Failure
-Decrease Fibrinogen
-Microcytosis.