Haematological and molecular characterisation of high haemoglobin F among anaemic patients in Hospital Universiti Sains Malaysia

Anaemia is a condition usually associated with variety of diseases. In normal adults, haemoglobin F (HbF) levels are usually less than 1.0%. There are several genetic loci that have significant influence on HbF levels. The aim of this study is to determine the association between elevated HbF lev...

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Main Author:	Ghani, Siti Nor Assyuhada Mat
Format:	Thesis
Language:	English
Published:	2020
Subjects:	R Medicine
Online Access:	http://eprints.usm.my/48920/1/SITI%20NOR%20ASSYUHADA%20BINTI%20MAT%20GHANI-FINAL%20THESIS%20P-SKM000518%28R%29%20PWD_-24%20pages.pdf
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spelling	my-usm-ep.489202021-04-19T02:37:17Z Haematological and molecular characterisation of high haemoglobin F among anaemic patients in Hospital Universiti Sains Malaysia 2020-12 Ghani, Siti Nor Assyuhada Mat R Medicine Anaemia is a condition usually associated with variety of diseases. In normal adults, haemoglobin F (HbF) levels are usually less than 1.0%. There are several genetic loci that have significant influence on HbF levels. The aim of this study is to determine the association between elevated HbF level with haematological parameters and the presence of the BCL11A (rs1186868) and HMIP-2 (rs9376090) SNPs in anaemic patients due to acquired causes. This study involved 144 anaemic patients from Hospital Universiti Sains Malaysia (HUSM) with HbF level > 1.0%. High-performance liquid chromatography (HPLC) was used to determine the HbF and HbA2 levels. Multiplex ARMS-PCR and gap-PCR were performed for those samples with high HbA2 level (>3.2%) and normal HbA2 level (≤3.2%) respectively to detect mutation at β-globin gene cluster. Allelic discrimination for rs1186868 and rs9376090 were performed using real-time PCR technique for samples with no mutation detected. In this study, the mean age of patients is 19.99 ± 1.64 years with female 61.1% predominance. Majority were Malays (99.3%). There was a moderate negative correlation and statistically significant between HbF level with Hb level and RBC count, (r = -0.348, P < 0.05) (r = -0.377, P < 0.05) respectively. Meanwhile, the correlation between HbF level with MCV and MCH showed weak negative correlation but not statistically significant, (r = -0.079, P > 0.05) (r = -0.073, P > 0.05) respectively. Following multiplex ARMS-PCR, 65 (74.7%) mutations were detected which comprises of 49 heterozygous Cd 26 (75.4%), 10 heterozygous Cd 41/42 (15.4%), 3 compound heterozygous Cd 26 and Cd 41/42 (4.6%) and 3 heterozygous IVS 1–1 (4.6%), while 22 patients were not detected. Out of 57 samples, only one patient (1.8%) was found positive with Thai (δβ)°-thalassaemia type deletions when subjected to the multiplex gap-PCR consisted of four target deletions; Siriraj J Gγ(Aγδβ)o-thal, Thai (δβ)°-thalassaemia, HPFH-6, and Hb Lepore. There was a significant difference between the mean of HbF level of patients with and without β-globin gene cluster mutation and deletion (P < 0.05). The minor allele frequency (MAF) in both rs1186868 and rs9376090 shows similar to the East Asian (EAS) population. There is no significant difference of HbF level between genotypes containing HbF-promoting alleles of rs9376090 (TC and CC) when compared to genotype TT (P > 0.05). In conclusion, HbF level correlates with anaemic status of the subjects. Elevated HbF levels showed associations with both inherited and acquired causes. Additionally, there is no relationship between the increased HbF levels with the presence of SNPs rs9376090-C among acquired anaemic patients. Despite the need for further research in this area, this study provides data that can be used as a guideline for better anaemia treatment and management. 2020-12 Thesis http://eprints.usm.my/48920/ http://eprints.usm.my/48920/1/SITI%20NOR%20ASSYUHADA%20BINTI%20MAT%20GHANI-FINAL%20THESIS%20P-SKM000518%28R%29%20PWD_-24%20pages.pdf application/pdf en public masters Universiti Sains Malaysia Pusat Pengajian Sains Kesihatan
institution	Universiti Sains Malaysia
collection	USM Institutional Repository
language	English
topic	R Medicine
spellingShingle	R Medicine Ghani, Siti Nor Assyuhada Mat Haematological and molecular characterisation of high haemoglobin F among anaemic patients in Hospital Universiti Sains Malaysia
description	Anaemia is a condition usually associated with variety of diseases. In normal adults, haemoglobin F (HbF) levels are usually less than 1.0%. There are several genetic loci that have significant influence on HbF levels. The aim of this study is to determine the association between elevated HbF level with haematological parameters and the presence of the BCL11A (rs1186868) and HMIP-2 (rs9376090) SNPs in anaemic patients due to acquired causes. This study involved 144 anaemic patients from Hospital Universiti Sains Malaysia (HUSM) with HbF level > 1.0%. High-performance liquid chromatography (HPLC) was used to determine the HbF and HbA2 levels. Multiplex ARMS-PCR and gap-PCR were performed for those samples with high HbA2 level (>3.2%) and normal HbA2 level (≤3.2%) respectively to detect mutation at β-globin gene cluster. Allelic discrimination for rs1186868 and rs9376090 were performed using real-time PCR technique for samples with no mutation detected. In this study, the mean age of patients is 19.99 ± 1.64 years with female 61.1% predominance. Majority were Malays (99.3%). There was a moderate negative correlation and statistically significant between HbF level with Hb level and RBC count, (r = -0.348, P < 0.05) (r = -0.377, P < 0.05) respectively. Meanwhile, the correlation between HbF level with MCV and MCH showed weak negative correlation but not statistically significant, (r = -0.079, P > 0.05) (r = -0.073, P > 0.05) respectively. Following multiplex ARMS-PCR, 65 (74.7%) mutations were detected which comprises of 49 heterozygous Cd 26 (75.4%), 10 heterozygous Cd 41/42 (15.4%), 3 compound heterozygous Cd 26 and Cd 41/42 (4.6%) and 3 heterozygous IVS 1–1 (4.6%), while 22 patients were not detected. Out of 57 samples, only one patient (1.8%) was found positive with Thai (δβ)°-thalassaemia type deletions when subjected to the multiplex gap-PCR consisted of four target deletions; Siriraj J Gγ(Aγδβ)o-thal, Thai (δβ)°-thalassaemia, HPFH-6, and Hb Lepore. There was a significant difference between the mean of HbF level of patients with and without β-globin gene cluster mutation and deletion (P < 0.05). The minor allele frequency (MAF) in both rs1186868 and rs9376090 shows similar to the East Asian (EAS) population. There is no significant difference of HbF level between genotypes containing HbF-promoting alleles of rs9376090 (TC and CC) when compared to genotype TT (P > 0.05). In conclusion, HbF level correlates with anaemic status of the subjects. Elevated HbF levels showed associations with both inherited and acquired causes. Additionally, there is no relationship between the increased HbF levels with the presence of SNPs rs9376090-C among acquired anaemic patients. Despite the need for further research in this area, this study provides data that can be used as a guideline for better anaemia treatment and management.
format	Thesis
qualification_level	Master's degree
author	Ghani, Siti Nor Assyuhada Mat
author_facet	Ghani, Siti Nor Assyuhada Mat
author_sort	Ghani, Siti Nor Assyuhada Mat
title	Haematological and molecular characterisation of high haemoglobin F among anaemic patients in Hospital Universiti Sains Malaysia
title_short	Haematological and molecular characterisation of high haemoglobin F among anaemic patients in Hospital Universiti Sains Malaysia
title_full	Haematological and molecular characterisation of high haemoglobin F among anaemic patients in Hospital Universiti Sains Malaysia
title_fullStr	Haematological and molecular characterisation of high haemoglobin F among anaemic patients in Hospital Universiti Sains Malaysia
title_full_unstemmed	Haematological and molecular characterisation of high haemoglobin F among anaemic patients in Hospital Universiti Sains Malaysia
title_sort	haematological and molecular characterisation of high haemoglobin f among anaemic patients in hospital universiti sains malaysia
granting_institution	Universiti Sains Malaysia
granting_department	Pusat Pengajian Sains Kesihatan
publishDate	2020
url	http://eprints.usm.my/48920/1/SITI%20NOR%20ASSYUHADA%20BINTI%20MAT%20GHANI-FINAL%20THESIS%20P-SKM000518%28R%29%20PWD_-24%20pages.pdf
_version_	1747821972923875328

Haematological and molecular characterisation of high haemoglobin F among anaemic patients in Hospital Universiti Sains Malaysia

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