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

Area of Science:

• Hematology
• Medical Diagnostics
• Genetics

Background:

• Thalassemia and thalassemic hemoglobinopathies are significant health concerns in India, causing substantial morbidity and mortality.
• The Indian population exhibits a wide array of hemoglobin variants due to its multiethnic nature.

Purpose of the Study:

• To analyze the laboratory aspects, including hematological profiles and HPLC findings, of detected hemoglobin variants.
• To discuss diagnostic challenges encountered in a routine clinical laboratory setting for hemoglobinopathies.

Main Methods:

• Screening of 4800 cases from a North Indian hospital-based population over two years.
• Utilized automated HPLC (Variant II Beta Thalassemia Short Program, Bio-Rad) for hemoglobin variant detection.
• Determined red cell indices using an automated hematology analyzer.

Main Results:

• Detected 290 cases with abnormal hemoglobin variants.
• Beta thalassemia was identified as the most prevalent variant, followed by hemoglobin E.
• Discussed diagnostic difficulties including borderline HbA2 values, silent mutations, alpha thalassemia, and rare variants.

Conclusions:

• Accurate laboratory diagnosis of hemoglobinopathies requires careful analysis of hematological parameters and HPLC results.
• Addressing diagnostic challenges, such as borderline HbA2 levels and coinheritance of beta thalassemia, is crucial for effective patient management.
• Correlation with genetic studies may be necessary for definitive diagnosis of complex or rare hemoglobin variants.