Role of HPLC in the detection of HbH disease

Wadhwa, Rachna; Singh, T. K.

doi:10.4103/0377-4929.81590

Cited by 3 publications

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“…It has been reported that one can detect HbH form of alpha thalassemia on HPLC by visual analysis of the chromatogram plot as it produces a sharp peak before the start of integration [ 29 ]. We, however, did not find any such case.…”

Section: Resultsmentioning

confidence: 99%

Detection of Abnormal Hemoglobin Variants by HPLC Method: Common Problems with Suggested Solutions

Pant

Kalita

Singh

et al. 2014

International Scholarly Research Notices

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Thalassemia and thalassemic hemoglobinopathies pose serious health problem leading to severe morbidity and mortality in Indian population. Plethora of hemoglobin variants is prevalent in multiethnic Indian population. The aim of the present study was to analyze laboratory aspects, namely, hematological profile and HPLC findings of the hemoglobin variants detected, and to discuss problems that we faced in diagnosis in a routine clinical laboratory. We screened a total of 4800 cases in a hospital based population of North India in a 2-years period of by automated HPLC method using the Variant Hemoglobin Testing System (Variant II Beta Thalassemia Short Program, Bio-Rad Laboratories) under the experimental conditions specified by the manufacturer. Whole blood in EDTA was used and red cell indices were determined using automated hematology analyzer. We detected 290 cases with abnormal variants in which beta thalassemia was the most common followed by hemoglobin E. Here, we discuss the laboratory aspects of various hemoglobin disorders and diagnostic difficulties in cases like borderline HbA2 values, presence of silent mutation, alpha thalassemia gene, and few rare variants which at times require correlation with genetic study. Special attention was given to HbA2 level even in presence of a structural variant to rule out coinheritance of beta thalassemia gene.

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Section: Resultsmentioning

confidence: 99%

Detection of Abnormal Hemoglobin Variants by HPLC Method: Common Problems with Suggested Solutions

Pant

Kalita

Singh

et al. 2014

International Scholarly Research Notices

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“…The diagnosis of Hb H disease in our case depended on the document of Hb H bands in the second and the third Hb analyses which were significantly sharp peak on the chromatogram by the HPLC method [9] and later it was confirmed by the genotype study and finally found positive for SEA and 3.7 kb deletions, the common genetic co-inheritance of Hb H disease [10]. Likewise, the iron deficiency anemia was diagnosed because the serum ferritin level was less than 30 ng/ml [11].…”

Section: Discussionmentioning

confidence: 84%

The absence of hemoglobin Hb H band in the proven hemoglobin H disease patient due to the iron deficiency anemia

Insiripong¹,

Insiripong²

2018

Blood Heart Circ

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Introduction: Iron deficiency anemia has not only been rarely seen among hemoglobin H disease patients but can mask the diagnosis of hemoglobin H disease also.Case Presentation: A 50-year-old Thai woman had marked and progressive anemia with just splenomegaly on the physical examination. Her blood tests showed: Hb 3.8 g%, MCV 50.4 fl, MCH 15.4 pg, ferritin 5.0 ng/ml, serum iron 33 ug/dl, TIBC 123 ug/dl, the Hb analysis using the high-performance liquid chromatography method found only A 2 A, Hb A 2 2.1 %. Iron deficiency anemia was diagnosed and continuously treated with the oral iron tablets. After two months of treatment, her blood showed: Hb 7.7 g%, MCV 69.3 fl, MCH 20.5 pg, reticulocyte 6.0 %, ferritin 6.2 ng/ml, Hb analysis showed: A 2 AH Bart, Hb A 2 1.7 %. The diagnosis of Hb H disease was added. Later Hb H was confirmed by the positive PCR for alpha thalassemia-1, Southeast Asian deletion and alpha thalassemia-2, 3.7 kb deletion, genes. After five months of the iron treatment, Hb 8.8 g%, MCV 59.3 fl, MCH 16.7 pg, ferritin 110.7 ng/ml, Hb analysis: A 2 ABartH, Hb A 2 1.4 %. The band of Hb H was found after Hb concentration was raised even though the ferritin level was still low. Conclusion:The absence or presence of Hb H band of the Hb H disease patient on Hb electrophoresis seemingly depended on the hemoglobin concentration more than the sufficiency of the iron storage.

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“…HbH disease shows considerable variability in clinical and haematological severity. [22] Due to socio-cultural practices, marriages in India are usually among individuals of the same caste or ethnic group and this makes it important to know the prevalence of different abnormal Hb variants in different ethnic groups. [23] In our country premarital screening is still considered taboo.…”

Section: Discussionmentioning

confidence: 99%

A Study of Haemoglobinopathies and Haemoglobin Variants Using High Performance Liquid Chromatography (Hplc) in a Teaching Hospital of Odisha

Raman¹,

Sahu²,

Senapati³

2017

jemds

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BACKGROUND Thalassaemias and haemoglobinopathies are highly prevalent in India. Identification of these disorders is important for epidemiologic purposes and for prevention of thalassaemia major and clinically severe haemoglobinopathies. The use of high performance liquid chromatography (HPLC) as a screening method for detection of these groups of disorder is increasing in last two decades. The aim is to study of thalassaemias and haemoglobinopathies using high performance liquid chromatography as a diagnostic tool in patients of a tertiary care hospital of Odisha. METHODS AND MATERIALS A total of 788 cases were included in the study. Samples were analysed on the BIO-RAD D-10 TM dual mode HPLC system. Personal history, family history, peripheral blood findings and sickling test result were correlated. Family studies were done whenever required and possible. However, secondary confirmatory tests were not done. Statistical analysis was done on Microsoft Excel. Continuous variables were expressed as mean ± SD. Categorical variables were expressed as frequencies and percentages. RESULTS Abnormal HPLC patterns were seen in 37.18% of cases. Sickle cell heterozygous (15.10%), sickle cell homozygous (9.90%) and βthalassaemia trait (6.10%) were the most common abnormalities found. Other patterns detected include β-thalassaemia major and intermedia, compound heterozygous state of HbS and β-thalassaemia, HbE trait, HbE disease, double heterozygous state of HbS and HbE, HbE and β-thalassaemia and HbS and hereditary persistent of foetal haemoglobin, HbD Punjab trait, Hb Lepore trait and HbH disease. CONCLUSION Sickle cell along with β-thalassaemias are the major abnormal haemoglobins in Odisha. Premarital and antenatal screenings are important measures to prevent birth of children with severe haemoglobin disorders. HPLC is a rapid and reliable technique for identification of various haemoglobin fractions.

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Role of HPLC in the detection of HbH disease

Cited by 3 publications

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Detection of Abnormal Hemoglobin Variants by HPLC Method: Common Problems with Suggested Solutions

Detection of Abnormal Hemoglobin Variants by HPLC Method: Common Problems with Suggested Solutions

The absence of hemoglobin Hb H band in the proven hemoglobin H disease patient due to the iron deficiency anemia

A Study of Haemoglobinopathies and Haemoglobin Variants Using High Performance Liquid Chromatography (Hplc) in a Teaching Hospital of Odisha

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