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| Year : 2010 | Volume
: 53
| Issue : 4 | Page : 861-862 |
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| High performance liquid chromatography as an aid in picking up hemoglobin H disease |
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Arpita R Dam, Ritesh Sachdev
Clinical Reference Lab, Super Religare Laboratories, New Delhi, India
Click here for correspondence address and email
| Date of Web Publication | 27-Oct-2010 |
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How to cite this article:
Dam AR, Sachdev R. High performance liquid chromatography as an aid in picking up hemoglobin H disease. Indian J Pathol Microbiol 2010;53:861-2 |
Sir,
Alpha thalassemias are a varied group of hemoglobinopathies with defective alpha chain synthesis. The alpha genes are duplicated and located in the telomeric region of chromosome 16. Hematological conditions encountered include silent carrier (single gene defect, -α/αα), alpha thalassemia trait (-α/-α or --/αα), hemoglobin (Hb) H disease ( --/-α), and Hb Barts fetalis( --/--).
Carriers and trait are best diagnosed by globin chain analysis and Hb Barts is usually incompatible with postnatal life.
We report a case of Hb H diagnosed with the help of high-pressure liquid chromatography (HPLC) and ancillary inputs. We received blood sample of a 13-year-old boy for Hb HPLC. Complete blood counts revealed a Hb of 5.2 g/dL, microcytic hypochromic (MCHC) blood picture [1] with marked anisopoikilocytosis, target cells, and schistocytes [Figure 1]a,b. The red cell indices were MCV 70.3 fL, MCH 19.3pg, Mean corpuscular hemoglobin concentration 27.5 g/dL and RDW 41.2%. Patient had hepatosplenomegaly and anemia, which was not attributed to any nutritional deficiency. Routine investigations for other causes of anemia were noncontributory. There was no history of blood transfusion. HPLC studies were advised to rule out any hemoglobinopathy. HPLC revealed a low Hb A2 of 1.7% along with pre run peaks (AHBarts pattern, with Hb Barts and H eluting as pre run peaks) before Hb F [Figure 1]c. Reticulocyte preparation with 1% brilliant cresyl blue showed reticulocytosis along with typical "golf ball" Hb H inclusions approximately 40% [Figure 2]. A provisional diagnosis of Hb H disease was made and globin chain analysis/DNA studies were recommended. | Figure 1: (a,b): Peripheral smear with microcytic hypochromic blood picture, anisopoikilocytosis with schistocytes, (c) HPLC showing prerun peaks of Hb Barts and Hb H along with decreased Hb A2
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 | Figure 2: Reticulocyte preparations with brilliant cresyl blue showing Hb H "golf ball" inclusions with typical reticulocytes
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Hb H [2] is usually due to deletion of three alpha genes (deletional Hb H). The rest are due to deletion of two genes along with point mutation of third gene (nondeletional Hb H). It is common in South East Asia. Due to decreased alpha chains and excess of β chains, tetramers of β chain (β4) are formed. Hb H being uns table precipitates in red cells, causing hemolysis. High oxygen affinity also contributes to the severity of the disease. Clinically it resembles β thalassemia intermedia.
High index of suspicion together with awareness of the condition can help in early pick up even on routine testing. The important aids in picking up the condition on HPLC include low Hb A2 (less than 2.5%) and prerun peaks before the point of integration, present before Hb F. These together with MCHC blood picture, target cells, schistocytes, and reticulocyte preparations showing typical inclusions help in diagnosing the condition with reasonable accuracy. They must be supplemented with molecular studies for confirmation.
 Acknowledgment | |  |
Sincere thanks to our Director, Dr Ashok Rattan, Dr Aparna Ahuja and Executive Director, Dr Amar Dasgupta for their constant support and guidance.
| References | | |
| 1. | Jolobe OM. Haemoglobin-H disease presenting with microcytic hypochromic anaemia in an 81 year old woman. Postgrad Med J 1993;69:930-1. 
[PUBMED] [FULLTEXT] |
| 2. | Chui DH. Alpha-thalassemia: Hb H disease and Hb Barts hydrops fetalis. Ann N Y Acad Sci 2005;1054:25-32.
[PUBMED] [FULLTEXT] |
Correspondence Address:
Ritesh Sachdev
A 803, Plot 7A, Navrattan Apartments, Sector 23, Dwarka, New Delhi 110075
India
 Source of Support: None, Conflict of Interest: None  | Check |
DOI: 10.4103/0377-4929.72008
[Figure 1], [Figure 2] |
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