open access

Vol 48, No 1 (2017)
Prace oryginalne / Original research articles
Published online: 2017-01-01
Submitted: 2016-03-08
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Diagnosing Beta Thalassemia trait in a developing country

Shan-e- Rauf, Ghassan Umair Shamshad1, Fareeha Mushtaq1, Saleem Ahmed Khan1, Nadir Ali1
DOI: 10.1016/j.achaem.2017.01.001
·
Acta Haematol Pol 2017;48(1):18-22.
Affiliations
  1. Armed Forces Institute of Pathology, Rawalpindi, Pakistan

open access

Vol 48, No 1 (2017)
Prace oryginalne / Original research articles
Published online: 2017-01-01
Submitted: 2016-03-08

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.

Get Citation

Keywords

Beta Thalassemia trait; HPLC; Cellulose Acetate electrophoresis; Polymerase Chain Reaction; HbA2

About this article
Title

Diagnosing Beta Thalassemia trait in a developing country

Journal

Acta Haematologica Polonica

Issue

Vol 48, No 1 (2017)

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

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