BETA CHAIN SINGLE AMINO ACID SUBSTITUTIONS
Hb SS Hb AS
Informations Sickle Cell Anemia
Predominant in Africans
Migrates slower than Hb A
Most Common Hemoglobin
Variant
Inheritance A2B2 6Glu to Val
Autosomal codominant gene
Homozygous Dominant
RBC appearance Severe Normocytic, Hypochromic
Anemia
Lab findings Anisocytosis
Poikilocytosis
Polychromasia
Reticulocytosis
Leukocytosis = Due to persistent
infections
Thrombocytosis = Due to fibrotic
spleen
Cell and variants present Sickle Cells = aka Drepanocytes
Target Cells, Ovalocytes and
Schistocytes (fragments of RBCs)
Basophilic Stipplings, Howell-
Jolly Bodies and Pappenheimer
Bodies
**Howell-Jolly and Pappenheimer Bodies
indicate NONFUNCTIONAL SPLEEN
(autosplenectomy).
Test Dithionite Tube Test / Sickle Cell
Solubility (+)
HbS Electrophoresis (+)
Osmotic Fragility Test (OFT) and
ESR Decreased
RBC Half-Life in SCA = only 10-20
Days
Myeloid:Erthyroid Ratio (M:E Ratio)
in SCA = 1:1
Increased LDH, Unconjugated
Bilirubin (B1), Urobilinogen
Decreased Haptoglobin
Treatment Cyanate and Nitrogen Mustard = In
Vitro Antisickling Agents
Bone Marrow Transplantation
Hb S-Thalassemia
Sickle Cell Trait
HbA (normal adult haemoglobin) is
present in higher concentration than
HbS
Usually no symptoms occur
Incidence is great in Africans and Black
Americans.
NO ANEMIA
A2B1B2 6Glu-Val
Heterozygous Dominant state of sickle
cell disease.
Normocytic, Normochromic
(low urine Hematuria and Hyposthenuria
concentration) may occur
Possible few target cells and occasional
sickle cell
Sickle Cell Solubility Test (+)
Hb Electrophoresis = Hb A and Hb S (+)
TOLEDO, ML
Hb CC Disease Hb AC
Most Common Sickle Cell Disease Crystal Hemoglobin Hemoglobin C Trait
2nd Most Common Hemoglobin Common in West
Variant in Mediterranean Populations
Variant after HbS Africans
Most Common in Black Races and No anemia
found mostly in Africa
Doubly heterozygous condition in which A2B2 6Glu-Lys A2B2,A2B2 6Glu-Lys
both HbS and thalassemia is inherited by Homozygous Dominant = HbCC Heterozygous Dominant
a single individual (Hemoglobin C Disease) for Hb C
Heterozygous Dominant = HbAC
(Hemoglobin C Trait)
Microcytic, Hypochromic RBCs Normocytic, Normochromic Anemia Normocytic, Slightly
Hypochromic RBCs
Anisocytosis (High RDW) Reticulocytosis occurs HbA is of higher
MCV and MCH are normal but concentration than HbC.
Retic Count = Normal or Slightly
MCHC is increased.
Decreased Decreased Red Cell Survival
Many Target Cells Numerous Target Cells (40-90%) Microspherocytes
and Few Spherocytes present and Increased
(Microspherocytes) Target Cells
HbC Crystals = Appear as
Hexagonal or Rod-Shaped Crystals
that are Blunt on Both Ends.
Sickle Cell Solubility Test or Dithionite Electrophoresis = Increased HbC
Tube Test (+) and HbF but Absent HbA
Electrophoresis = Lesser HbA than Citrate Agar Electrophoresis helps
HbS separate HbC from other
haemoglobin variants.
Increased HbA2 and HbF
Column Chromatography =
Beta to Alpha Chain Ratio in HbS-Beta Separates and Quantitates HbC and
Thalassemia = 0.5:1 HbA2 when both are present in a
specimen.
B-A Chain Ratio in SS or AS = 1:1
(Normal)
Mainly Supportive Patients are generally asymptomatic No symptoms
Transfusion causes temporary and no treatment is required.
dilutional effects and reduces HbS,
HbF and HbA2 levels on
electrophoresis
 BETA CHAIN SINGLE AMINO ACID SUBSTITUTIONS BETA CHAIN DOUBLE AMINO ACID
SUBSTITUTIONS
Hb SC Disease Hb D Hb SD Hb E Hb O Arab Hb C Harlem

Informations Milder than Hb SS and fewer Migrates at the same position as Only HbSD-Los 3rd Most Common Found in Middle HbC-Georgetown
complications Hb S in alkaline pH but does not Angeles produces a Hemoglobin Variant after Eastern countries.
cause Sickling clinical abnormality HbS and HbC
Most Common HbD Variant = similar to that of mild Greatest frequency in
HbD-Los Angeles (aka HbD- sickle cell anemia Southeast Asia
Punjab) HbE + Beta-Thalassemia =
Found principally in Pakistan Condition Similar to Beta-
and Northwest India. Thalassemia Major
Most common HbD Variant in
the US.

Inheritance A2B2 6Glu-Val and A2B2 A2B2 121Glu-Gln A2B2 6Glu-Val and A2B2 26Glu-Lys A2B2 121Glu-Lys A2B2 6Glu-Val and A2B2 73Asp-
6Glu-Lys Homozygous Dominant = HbDD A2B2 121Glu-Glycine Homozygous D = Hb EE Asn
Doubly heterozygous Heterozygous Dominant = Doubly heterozygous Heterozygous D = HbAE
Codominant inheritance HbAD (A2B2, A2B2 121Glu- (A2B2, A2B2 26Glu-Lys)
pattern Gln)

RBC appearance Mild Normocytic, Normal RBC Indices Homozygous Hb EE:

Normochromic Anemia Splenomegaly.
Clin/Lab findings Splenomegaly Homozygous HbD = Mild Microcytic, Homozygous HbO-
Aseptic Necrosis on Femoral Splenomegaly and Mild Normochromic Hemolytic Arab = Splenomegaly
Head Hemolytic Anemia Anemia Mild Hemolytic Anemia
Retinal Diseases Heterozygous HbD = No Clinical Red Cell Survival Time = w/ numerous target cells.
Abnormalities Slighty Decreased Heterozygous HbO-
Numerous Target Cells Arab = Asymptomatic
Heterozygous Hb AE: HbO-Arab + HbS =
Elevated RBC Count Severe Condition
Increased Target Cells similar to SCA
Normal Red Cell Survival
Patients are not Anemic

Cell and variants Target Cells present and Numerous Flat Target Cells
present Characteristic Pocketbook
Cells
HbSC Crystals =
Characteristic Washington
Monument Appearance
(Fingerlike Projections that
Protrude the Cell Membrane)
Test Sickle Cell Solubility Test (+) Sickle Solubility Test (-) Most Definitive Test of Electrophoresis = HbE, HbC, Migrates together with Sickle Cell Solubility (+)
Electrophoresis = Equal Electrophoresis HbSD = Citrate Agar HbO-Arab and HbA2 HbA2, HbC and HbE on Electrophoresis = Band in HbC
Amounts of HbS and HbC, Migrates on Same Position as Electrophoresis migrates closely on Cellulose Cellulose Acetate position on Cellulose Acetate.
HbF Present or Increased HbS on Cellulose Acetate. (separates HbSS from Acetate. Citrate Agar E =
and no HbA Migrates on Same Position as SD) HbE migrates slightly faster Required for
HbA and HbA2 on Citrate Agar. than HbC and HbA2 on Differential Diagnosis.
Sickle Cell Solubility (+) Cellulose Acetate HbO-Arab moves just
Citrate Agar Electrophoresis slightly away from point
= Differentiates HbC from of application to cathode
HbO-Arab
Treatment Therapy TOLEDO, ML
Sodium Bicarbonate
Magnesium Sulfate
Heparin
 ALPHA CHAIN
SUBSTITUTIONS
HbG-Philadelphia
Informations Only Alpha Chain Variant
of Significance in the US
Most Common Variant
among American Blacks
after HbS and HbC
OTHER HEMOGLOBIN VARIANTS GLOBIN CHAIN AMINO GLOBIN CHAIN ELONGATION
ACID DELETION
Hb I Hb Barts and Hb H Hb Gun Hill Hb Constant Spring (Hb Hb M
CS)
Affects either Alpha or Two variants found in Results to Altered Resembles Alpha- Associated with Methemoglobin and
Beta Chain at Various patients with Alpha- Hemoglobin Structure that Thalassemia Congenital Cyanosis
Positions Thalassemia NO HEME CAN BIND TO Prevalent among Greeks Prevalent in Japanese
THE BETA CHAIN and Chinese
Due to 2 Beta Chains of
UNEQUAL LENGTH

Inheritance A2 (68Asn-Lys) B2 Glutamic Acid replaces Hb Barts = Occurs in
Lysine at different Newborn Infants and
positions consists of 4 Gamma
Chains.
Hb H = Occurs in Older
Children and consists of 4
Normal Beta Chains
RBC appearance
Deletion of 5 Beta-Chain A2 (141+32) = A2(172)
AA (91st to 95th AA) =
B91-95
Hb M-Boston = A2 (58Tyrosine) B2
Elongated Alpha Chain Hb M-Iwate = A2 (87Tyr) B2
due to Addition of 31 AA Hb M-Saskatoon = A2B2 (63Tyr)
to end of chain Hb M-Hyde Park = A2B2 (92Tyr)
Often inherited as a Hb M-Milwaukee = A2B2 (67Glutamic
Double Heterozygous Acid)
condition = Hb H
Constant Spring
Homozygous Hb CS =
Mild Microcytic, Hemolytic
Anemia

Heterozygous Hb CS =
Little or No Anemia,
Microcytic Cells w/ Target
Cells

Clin/Lab findings Results to Unstable Lavender Blue (Cyanotic) Skin Color

Test Migrates similar to HbS
on Cellulose Acetate.
Sickle Cell Solubility (-)
Migrates similar to HbA
on Citrate Agar.
Abnormal Alpha Chain
moves farther toward
cathode
Abnormal Hb A2 = Aka
HbG2 = Migrates as
Minor, Faint Band toward
Cathode (opposite from
point of application).
I Counterstaining
procedure
TOLEDO, ML
Hemoglobin and Clubbed Fingers
produces Mild Hemolytic Chocolate Brown Blood
Anemia. Causes Globin Chain Precipitation
Heinz Bodies present
Migrates between Hb Both migrates farther Cellulose Acetate = Cellulose Acetate Diagnosis = Electrophoresis @ pH 7.1
Barts and Hb H on toward the anode than Migrates close to Hb A2 Electrophoresis = Add Potassium Cyanide before
Cellulose Acetate HbA Migrates slower than Hb electrophoresis to oxidize all
Citrate Agar = Migrates A2. haemoglobin to methemoglobin
Hb H and Hb I = Best with Hb A or Between
Differentiated by Hb A and Hb S Must be counterstained Spectrophotometric Analysis =
Cellulose Acetate @ pH with Ortho-Toluidine or Identification of Hb M Variants (each has
7.0 Ortho-Dianisidine. unique absorption range)
o Hb H migrates
but Hb I does Differentiates it from Confirmation = Amino Acid Chain
not nonheme-protein. Studies
Hb CS = Purple
Nonheme-Protein = Pink