The significance of capillary electrophoresis in the detection of haemoglobin constant spring and its relationship with high-performance liquid chromatography

Introduction: Haemoglobin Constant Spring (Hb CS) is one of the most common non-deletional types of alpha (α) thalassaemia in Southeast Asia region. The nature of this abnormal globin gene is that it is unstable, labile and is present in minute amount in the peripheral blood. Thus, this may lead to...

Full description

Saved in:

Bibliographic Details
Main Author:	Rameli, Nabilah
Format:	Thesis
Language:	English
Published:	2021
Subjects:	TP200-248 Chemicals: Manufacture use etc.
Online Access:	http://eprints.usm.my/58926/1/NABILAH%20BINTI%20RAMELI-24%20pages.pdf
Tags:	Add Tag No Tags, Be the first to tag this record!

id	my-usm-ep.58926
record_format	uketd_dc
spelling	my-usm-ep.589262023-08-06T08:12:14Z The significance of capillary electrophoresis in the detection of haemoglobin constant spring and its relationship with high-performance liquid chromatography 2021 Rameli, Nabilah TP200-248 Chemicals: Manufacture, use, etc. Introduction: Haemoglobin Constant Spring (Hb CS) is one of the most common non-deletional types of alpha (α) thalassaemia in Southeast Asia region. The nature of this abnormal globin gene is that it is unstable, labile and is present in minute amount in the peripheral blood. Thus, this may lead to underdiagnosis of the disease. This study was conducted to determine the proportion of Hb CS among the Kelantan population and to compare range of peak value in Zone 2 CE findings for 3 groups of Hb CS (heterozygous, homozygous, and compound heterozygous) and their haematological parameters. The study aimed to look at the findings of HPLC in relation to CE results in detecting Hb CS. Study design and methods: This was a cross-sectional study involving secondary data collection from 378 samples which showed peak value on Zone 2 of CE. The samples were taken from the National Thalassemia screening of Form 4 students from all districts in Kelantan. The haematological parameters of red cells were analysed using Sysmex XN 3000 automated blood cell analyser, Hb analysis was performed using automated CE system (CAPILLARYS2 Flex-Piercing System Sebia), HPLC Biorad variant II, DNA analysed using multiplex polymerase chain reaction (PCR) and multiplex Amplification refractory mutation system (ARMS) to detect both deletional and non-deletional α-thalassaemia. Results: 376 samples (99.5%) with presence of peak value on Zone 2 of CE were confirmed to have termination codon CS mutation. Heterozygous Hb CS is the most common type of Hb CS detected in 344 samples (91.5%), followed by compound heterozygous Hb CS which was 31 samples (8.2%) and only 1 sample (0.3%) of homozygous Hb CS. The mean ± SD of peak value in Zone 2 of heterozygous Hb CS and compound heterozygous Hb CS were 0.61 ± 0.13 and 0.77 ± 0.34 respectively. The only sample of homozygous Hb CS showed the value of 4.9% of peak value in Zone 2 of CE. The significant differences of haematological parameters between heterozygous and compound heterozygous Hb CS were observed in haemoglobin level, MCV, MCH and MCHC. This study showed there was a good linear correlation between peak in C-window on HPLC and peak value in Zone 2 of CE in detecting Hb CS, r=0.73. Conclusion: Thus, by combining the haematological parameters and complementary tests of both CE and HPLC, the diagnosis of Hb CS can be detected prior to confirmation by DNA molecular study that is far more expensive. 2021 Thesis http://eprints.usm.my/58926/ http://eprints.usm.my/58926/1/NABILAH%20BINTI%20RAMELI-24%20pages.pdf application/pdf en public masters Universiti Sains Malaysia Pusat Pengajian Sains Perubatan
institution	Universiti Sains Malaysia
collection	USM Institutional Repository
language	English
topic	TP200-248 Chemicals: Manufacture use etc.
spellingShingle	TP200-248 Chemicals: Manufacture use etc. Rameli, Nabilah The significance of capillary electrophoresis in the detection of haemoglobin constant spring and its relationship with high-performance liquid chromatography
description	Introduction: Haemoglobin Constant Spring (Hb CS) is one of the most common non-deletional types of alpha (α) thalassaemia in Southeast Asia region. The nature of this abnormal globin gene is that it is unstable, labile and is present in minute amount in the peripheral blood. Thus, this may lead to underdiagnosis of the disease. This study was conducted to determine the proportion of Hb CS among the Kelantan population and to compare range of peak value in Zone 2 CE findings for 3 groups of Hb CS (heterozygous, homozygous, and compound heterozygous) and their haematological parameters. The study aimed to look at the findings of HPLC in relation to CE results in detecting Hb CS. Study design and methods: This was a cross-sectional study involving secondary data collection from 378 samples which showed peak value on Zone 2 of CE. The samples were taken from the National Thalassemia screening of Form 4 students from all districts in Kelantan. The haematological parameters of red cells were analysed using Sysmex XN 3000 automated blood cell analyser, Hb analysis was performed using automated CE system (CAPILLARYS2 Flex-Piercing System Sebia), HPLC Biorad variant II, DNA analysed using multiplex polymerase chain reaction (PCR) and multiplex Amplification refractory mutation system (ARMS) to detect both deletional and non-deletional α-thalassaemia. Results: 376 samples (99.5%) with presence of peak value on Zone 2 of CE were confirmed to have termination codon CS mutation. Heterozygous Hb CS is the most common type of Hb CS detected in 344 samples (91.5%), followed by compound heterozygous Hb CS which was 31 samples (8.2%) and only 1 sample (0.3%) of homozygous Hb CS. The mean ± SD of peak value in Zone 2 of heterozygous Hb CS and compound heterozygous Hb CS were 0.61 ± 0.13 and 0.77 ± 0.34 respectively. The only sample of homozygous Hb CS showed the value of 4.9% of peak value in Zone 2 of CE. The significant differences of haematological parameters between heterozygous and compound heterozygous Hb CS were observed in haemoglobin level, MCV, MCH and MCHC. This study showed there was a good linear correlation between peak in C-window on HPLC and peak value in Zone 2 of CE in detecting Hb CS, r=0.73. Conclusion: Thus, by combining the haematological parameters and complementary tests of both CE and HPLC, the diagnosis of Hb CS can be detected prior to confirmation by DNA molecular study that is far more expensive.
format	Thesis
qualification_level	Master's degree
author	Rameli, Nabilah
author_facet	Rameli, Nabilah
author_sort	Rameli, Nabilah
title	The significance of capillary electrophoresis in the detection of haemoglobin constant spring and its relationship with high-performance liquid chromatography
title_short	The significance of capillary electrophoresis in the detection of haemoglobin constant spring and its relationship with high-performance liquid chromatography
title_full	The significance of capillary electrophoresis in the detection of haemoglobin constant spring and its relationship with high-performance liquid chromatography
title_fullStr	The significance of capillary electrophoresis in the detection of haemoglobin constant spring and its relationship with high-performance liquid chromatography
title_full_unstemmed	The significance of capillary electrophoresis in the detection of haemoglobin constant spring and its relationship with high-performance liquid chromatography
title_sort	significance of capillary electrophoresis in the detection of haemoglobin constant spring and its relationship with high-performance liquid chromatography
granting_institution	Universiti Sains Malaysia
granting_department	Pusat Pengajian Sains Perubatan
publishDate	2021
url	http://eprints.usm.my/58926/1/NABILAH%20BINTI%20RAMELI-24%20pages.pdf
_version_	1776101245052780544

The significance of capillary electrophoresis in the detection of haemoglobin constant spring and its relationship with high-performance liquid chromatography

Similar Items