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Diagnosing Beta Thalassemia trait in a developing country
Affiliations- Armed Forces Institute of Pathology, Rawalpindi, Pakistan
open access
Abstract
Background
Beta Thalassemia trait (BTT) is diagnosed by detecting hemoglobin A2 (HbA2) >3.8% on either High Performance Liquid Chromatography (HPLC) or cellulose acetate electrophoresis (CAE). HPLC is an accurate and reproducible but costly alternative to more conventional CAE which is labor intensive but easy to interpret and inexpensive.
Objective
To determine the sensitivity of CAE and HPLC keeping PCR as gold standard for the diagnosis of BTT.
Study Design
Cross sectional.
Place and Duration of Study
Armed Forces Institute of Pathology Rawalpindi. May 2014 to January 2015.
Patient and Methods
Fiveml EDTA anti-coagulated blood was collected from 100 PCR proven cases of BTT. HbA2 levels were measured by running samples directly on HPLC. But for CAE, first a hemolysate was prepared which was then applied to cellulose acetate membrane at an alkaline pH (7.9). After elution of HbA2 band in Tris EDTA borate buffer (pH of 8.9), HbA2 concentration was calculated by measuring its absorbance in a photometer at a wavelength of 416nm.
Results
Mean age of the patients was 28.8±8.1 year. The most common mutation was Fr 8–9 (35%) followed by IVS1-5 (25%) mutation. Mean HbA2 levels by CAE and HPLC were 4.97±0.42 and 5.54±0.59 respectively. All the patients had HbA2>4% on both CAE and HPLC. None of our patients had false negative result either on CAE or HPLC.
Conclusion
CAE has comparable sensitivity with HPLC for detection of Beta Thalassemia Trait.
Abstract
Background
Beta Thalassemia trait (BTT) is diagnosed by detecting hemoglobin A2 (HbA2) >3.8% on either High Performance Liquid Chromatography (HPLC) or cellulose acetate electrophoresis (CAE). HPLC is an accurate and reproducible but costly alternative to more conventional CAE which is labor intensive but easy to interpret and inexpensive.
Objective
To determine the sensitivity of CAE and HPLC keeping PCR as gold standard for the diagnosis of BTT.
Study Design
Cross sectional.
Place and Duration of Study
Armed Forces Institute of Pathology Rawalpindi. May 2014 to January 2015.
Patient and Methods
Fiveml EDTA anti-coagulated blood was collected from 100 PCR proven cases of BTT. HbA2 levels were measured by running samples directly on HPLC. But for CAE, first a hemolysate was prepared which was then applied to cellulose acetate membrane at an alkaline pH (7.9). After elution of HbA2 band in Tris EDTA borate buffer (pH of 8.9), HbA2 concentration was calculated by measuring its absorbance in a photometer at a wavelength of 416nm.
Results
Mean age of the patients was 28.8±8.1 year. The most common mutation was Fr 8–9 (35%) followed by IVS1-5 (25%) mutation. Mean HbA2 levels by CAE and HPLC were 4.97±0.42 and 5.54±0.59 respectively. All the patients had HbA2>4% on both CAE and HPLC. None of our patients had false negative result either on CAE or HPLC.
Conclusion
CAE has comparable sensitivity with HPLC for detection of Beta Thalassemia Trait.
Keywords
Beta Thalassemia trait; HPLC; Cellulose Acetate electrophoresis; Polymerase Chain Reaction; HbA2
About this articleTitle
Diagnosing Beta Thalassemia trait in a developing country
Journal
Issue
Pages
18-22
Published online
2017-01-01
DOI
10.1016/j.achaem.2017.01.001
Bibliographic record
Acta Haematol Pol 2017;48(1):18-22.
Keywords
Beta Thalassemia trait
HPLC
Cellulose Acetate electrophoresis
Polymerase Chain Reaction
HbA2
Authors
Shan-e- Rauf
Ghassan Umair Shamshad
Fareeha Mushtaq
Saleem Ahmed Khan
Nadir Ali