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HAEMOGLOBINOPATHY CASES on HPLC Dr. Archana Vazifdar Head Pathologist Hindlabs, HLL Lifecare Ltd.
HPLC Automated system precalibrated column and gradient Direction of flow Detector
Hemoglobin is eluted in a stepped manner by Buffers of Increasing Ionic strength
CHROMATOGRAMSOutputTime Peak RT RT (min) Area Total Area Count: 1-3 million HbF: 1-2%RT: HbA2: % (2-3.6%)RT:
P2- Glycemic status, upto 6% acceptable P3- 6% acceptable, 6-15% deterioration 15-25% HbJ
8 mnth/M, pallor, failure to thrive
Homozygous beta thalassemia
Heterozygous Beta thalassemia Mother of 8 mnth old child
HbA2- Normal RBC indices- Normal – Silent β thalassaemia RBC indices s/o thal – Co existing IDA – Co inheritance of α thalassaemia – δβ thalassaemia HbA2 LOW in α thalassaemia
HbA2% HbF% ≤ 2 Hb N MCV N Symptoms - FINAL NORMAL 4-10≤ 2/ mBorderline Asymptoma tic β THAL TRAIT NN Asymptoma tic Hb LEPORE TRAIT 25-35≤ 1NN Asymptoma tic HbE TRAIT 40-48≤ 2NN Asymptoma tic HbD IRAN Heterozygo us 50-60≥ 10Severe HbE/Hbβ double heterozygou s ≥ mAsymptoma tic HbE homozygou s
Hb Lepore Trait
Heterozygous E thalassemia
All criteria fit provided there is NO history of recent blood transfusion ALWAYS CORRELATE with clinical history with CBC & peripheral blood picture Degenerated sample
Hb 7 MCV 84 MCH 28 MCHC 32.4 RDW 20.2 Case 1: 2 yr/M, anemia, hepatosplenomegaly
Elevated HbF: HPFH (HbF 5-30%, CBC N, asymp) Heterozygous δβ thal (HbF 3-20%, asymp) Homozygous beta thal Pregnancy
Recently transfused c/o homozygous beta thalassemia
Case 2: 28/F, Severe anemia
Borderline HbA2 levels: Silent carrier of thalassemia β thal with superadded IDA Macrocytosis αβ thalassemia
Borderline elevated HbA2 due to macrocytosis
P3: 6% acceptable, 6-12% deterioration 15-25% HbJ
Hb: 6.5 MCV: 63 MCH: 23 MCHC: 26 RDW: 19.6
Double heterozygous for HbE and β thalassaemia
Tests may not be accurate if… Patient had a blood transfusion within the past four months. Patient has polycythemia (increased red blood cell production) or underlying anemia If the patient is on certain medications Aged/ degenerated sample
THALASSAEMIA A group of chronic, inherited anemias characterised by defective Hemoglobin (Hb) synthesis and ineffective erythropoiesis, particularly common.
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CORE AREA 4 HAEMATOLOGY GROUP C. Full Blood Count-Case C A 25 year old female: A 25 year old female: anaemia anaemia Never pregnant Never pregnant No.
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$100 $200 $300 $400 $500 $100$100$100 $200 $300 $400 $500 Topic 1Topic 2Topic 3Topic 4 Topic 5 FINAL ROUND.
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