HEMATOLOGY: RBC THALASSEMIA

Romie Solacito, MLS4C

Globin Synthesis Abnormalities
 Thalassemia – quantitative abnormalities
 Hemoglobinopathies – qualitative abnormalities
o Defects on the Amino Acid present:
Amino Acid Substitution and Amino Acid
Elongation

Genetics of Globin Synthesis

 Chromosome 16: Alpha gene and Zeta gene
 Chromosome 11: Beta gene, Delta gene, Gamma
gene, and Epsilon gene

Six Normal Hemoglobin

 Embryonic Hemoglobin:
o Gower 1 – Zeta and Epsilon
o Gower 2 – Alpha and Epsilon
o Portland – Zeta and Gamma
 Fetal Hemoglobin:
o Hemoglobin F – Alpha and Gamma
 Adult Hemoglobin  1st 2 months of gestation – Gower 1 (zeta &
o A1 – Alpha and Beta epsilon)
o A2 – Alpha and Delta  2nd month of gestation – Gower 2 (alpha and
epsilon); High Portland (zeta and gamma)
Development Expression
  10 weeks of gestation – switching (zeta to alpha)
at C16; switching (epsilon to gamma) at C11;
formation of Hemoglobin F (alpha and gamma)
 Throughout Fetal Life – Hemoglobin F
 Shortly After Birth – DOWN regulation of gamma
chain; UP regulation of beta chain; Hemoglobin
A (alpha and beta)
 6 months of Adulthood – Hemoglobin A1 (alpha
and beta); low amount of A2 (alpha and delta)
Thalassemia: Genetic Defects
 Reduced or absent transcription of mRNA
o Decreased RNA stability
 mRNA processing error
o results to no globin chain or altered
globin chain production
 Translation error
o Frameshift Mutation
o Missense Mutation
o Nonsense Mutations
 Deletion of one or more globin chains
Pathophysiology
 Reduced or absent production of a particular
globin chain with reduced hemoglobin synthesis
 Unequal production of the alpha or beta chain
Beta-Thalassemia Mechanism
 Asymptomatic till 6 months due to increase
Hemoglobin F
 Symptoms appear between 6 to 24 months after
the compensation of gamma to beta switching
 Increased Erythropoietin but still exhibits
ineffective erythropoiesis; promotes Bone
Marrow Expansion
 Increased Iron accumulation: increased
Erythropoietin = decreased hepcidin
 Progressed to reduction of bone mineral
thinning
o Increased risk of pathologic fractures
 o Exhibit frontal bossing PARAMETERS THALASSEMIA IRON DEFICIENCY
Blood Picture Micro, Hypo Micro, Hypo
Alpha-Thalassemia Mechanism Serum Ferritin ^/N v
 Decreased production of alpha chain Serum Iron N v
 Leads to the formation of excess gamma chain: TIBC v/N ^
Hemoglobin Barts (four gamma) from Transferrin v/N ^
Hemoglobin F
 After 6 months gamma switches to beta chain:
Hemoglobin H (four beta) from Hemoglobin A
 Hemoglobin Bart and Hemoglobin H
o Increased affinity to oxygen
o Causes marked hypoxia leading to
cardiac failure and Hydrops Fetalis
GENOTYPE DISORDER
--/-- Barts Hydrops Fetalis/α Thalassemia
Major
--/-α Hemoglobin H Disease
--/αα α Thalassemia Minor
α-/α- α Thalassemia Minor
αα/α- Silent Carrier
αα/αα None

Laboratory Diagnosis
 CBC with peripheral blood film: Microcytic,
Hypochromic
 Hematocrit: decreased (<35%)
 Hemoglobin: decreased (<12g/dL)
 RDW: increased (>14.5%)
 Electrophoresis:
o Cellulose Acetate – pH 8.4 to 8.6
 CSFA
o Citrate Agar – pH 6.0 to 6.4
 CSAF
 Cation-exchange HPLC
 Capillary Zone Electrophoresis
 Alkaline Denaturation Test
 Kleihauer – Betke Test
 Molecular Testing: Polymerase Chain Reaction

Fast Moving: Hemoglobin Bart and Hemoglobin H

CATHODE ANODE
C, G, E, As > S, G, D, Leopre > F > A > Portland > Barts > H