HAEMOGLOBINOPATHY CASES on HPLC Dr. Archana Vazifdar Head Pathologist Hindlabs, HLL Lifecare Ltd.
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Transcript of HAEMOGLOBINOPATHY CASES on HPLC Dr. Archana Vazifdar Head Pathologist Hindlabs, HLL Lifecare Ltd.
HAEMOGLOBINOPATHY CASESon HPLC
Dr. Archana VazifdarHead 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
CHROMATOGRAMS
Output
Time
Peak
RT (min)
Area
Total Area Count: 1-3 million
HbF: 1-2% RT: 1.03-1.13HbA2: 1.75-3.25% (2-3.6%) RT: 3.63-3.64
P2- Glycemic status, upto 6% acceptableP3- 6% acceptable, 6-15% deterioration
15-25% HbJ
8 mnth/M, pallor, failure to thrive
Homozygous beta thalassemia
22/Female
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%
2-3.6
HbF%
≤ 2
Hb
N
MCV
N
Symptoms
-
FINAL
NORMAL
4-10 ≤ 2/ m Borderline
Asymptomatic
β THAL TRAIT
10-18 2- 10 N N Asymptomatic
Hb LEPORE TRAIT
25-35 ≤ 1 N N Asymptomatic
HbE TRAIT
40-48 ≤ 2 N N Asymptomatic
HbD IRAN
Heterozygous
50-60 ≥ 10 SevereHbE/Hbβ double
heterozygous
≥ 60 2-10 m Asymptomatic
HbE homozygo
us
Hb Lepore Trait
34/M, Kolkata
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 7MCV 84 MCH 28MCHC 32.4RDW 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% deterioration15-25% HbJ
Hb: 6.5MCV: 63MCH: 23MCHC: 26RDW: 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