Detection of α-thalassaemia in neonates on cord blood and dried blood spot samples by capillary electrophoresis

Hafiza Alauddin, Mustafa Langa, Malisa Mohd Yusoff, Raja Zahratul Azma Raja Sabudin, Azlin Ithnin, Noor Farisah Abdul Razak, Nor Hidayati Sardi, Noor Hamidah Hussin

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Introduction: Haemoglobin Bart’s (Hb Bart’s) level is associated with α-thalassaemia traits in neonates, enabling early diagnosis of α-thalassaemia. The study aimed to detect and quantify the Hb Bart’s using Cord Blood (CB) and CE Neonat Fast Hb (NF) progammes on fresh and dried blood spot (DBS) specimen respectively by capillary electrophoresis (CE). Methods: Capillarys Hemoglobin (E) Kit (for CB) and Capillarys Neonat Hb Kit (for NF) were used to detect and quantify Hb Bart’s by CE in fresh cord blood and dried blood spot (DBS) specimens respectively. High performance liquid chromatography (HPLC) using the β-Thal Short Programme was also performed concurrently with CE analysis. Confirmation was obtained by multiplex ARMS Gap PCR. Results: This study was performed on 600 neonates. 32/600 (5.3%) samples showed presence of Hb Bart’s peak using the NF programme while 33/600 (5.5%) were positive with CB programme and HPLC methods. The range of Hb Bart’s using NF programme and CB programme were (0.5–4.1%) and (0.5-7.1%), respectively. Molecular analysis confirmed all positive samples possessed α-thalassaemia genetic mutations, with 23/33 cases being αα/--SEA, four -α3.7/-α3.7, two αα/-α3.7and three αα/ααCS. Fifty Hb Bart’s negative samples were randomly tested for α-genotypes, three were also found to be positive for α-globin gene mutations. Thus, resulting in sensitivity of 91.7% and 88.9% and specificity of 100% for the Capillarys Cord Blood programme and Capillarys Neonat Fast programme respectively. Conclusion: Both CE programmes using fresh or dried cord blood were useful as a screening tool for α-thalassaemia in newborns. All methods show the same specificity (100%) with variable, but acceptable sensitivities in the detection of Hb Bart.

Original languageEnglish
Pages (from-to)17-23
Number of pages7
JournalMalaysian Journal of Pathology
Volume39
Issue number1
Publication statusPublished - 1 Apr 2017

Fingerprint

Thalassemia
Capillary Electrophoresis
Fetal Blood
Hemoglobin E
High Pressure Liquid Chromatography
Mutation
Globins
hemoglobin Bart's
Early Diagnosis
Genotype
Polymerase Chain Reaction
Genes

Keywords

  • Capillary electrophoresis
  • Cord blood
  • Neonatal α-thalassaemia screening
  • α-thalassaemia

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Histology
  • Cell Biology

Cite this

Detection of α-thalassaemia in neonates on cord blood and dried blood spot samples by capillary electrophoresis. / Alauddin, Hafiza; Langa, Mustafa; Mohd Yusoff, Malisa; Raja Sabudin, Raja Zahratul Azma; Ithnin, Azlin; Abdul Razak, Noor Farisah; Sardi, Nor Hidayati; Hussin, Noor Hamidah.

In: Malaysian Journal of Pathology, Vol. 39, No. 1, 01.04.2017, p. 17-23.

Research output: Contribution to journalArticle

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abstract = "Introduction: Haemoglobin Bart’s (Hb Bart’s) level is associated with α-thalassaemia traits in neonates, enabling early diagnosis of α-thalassaemia. The study aimed to detect and quantify the Hb Bart’s using Cord Blood (CB) and CE Neonat Fast Hb (NF) progammes on fresh and dried blood spot (DBS) specimen respectively by capillary electrophoresis (CE). Methods: Capillarys Hemoglobin (E) Kit (for CB) and Capillarys Neonat Hb Kit (for NF) were used to detect and quantify Hb Bart’s by CE in fresh cord blood and dried blood spot (DBS) specimens respectively. High performance liquid chromatography (HPLC) using the β-Thal Short Programme was also performed concurrently with CE analysis. Confirmation was obtained by multiplex ARMS Gap PCR. Results: This study was performed on 600 neonates. 32/600 (5.3{\%}) samples showed presence of Hb Bart’s peak using the NF programme while 33/600 (5.5{\%}) were positive with CB programme and HPLC methods. The range of Hb Bart’s using NF programme and CB programme were (0.5–4.1{\%}) and (0.5-7.1{\%}), respectively. Molecular analysis confirmed all positive samples possessed α-thalassaemia genetic mutations, with 23/33 cases being αα/--SEA, four -α3.7/-α3.7, two αα/-α3.7and three αα/ααCS. Fifty Hb Bart’s negative samples were randomly tested for α-genotypes, three were also found to be positive for α-globin gene mutations. Thus, resulting in sensitivity of 91.7{\%} and 88.9{\%} and specificity of 100{\%} for the Capillarys Cord Blood programme and Capillarys Neonat Fast programme respectively. Conclusion: Both CE programmes using fresh or dried cord blood were useful as a screening tool for α-thalassaemia in newborns. All methods show the same specificity (100{\%}) with variable, but acceptable sensitivities in the detection of Hb Bart.",
keywords = "Capillary electrophoresis, Cord blood, Neonatal α-thalassaemia screening, α-thalassaemia",
author = "Hafiza Alauddin and Mustafa Langa and {Mohd Yusoff}, Malisa and {Raja Sabudin}, {Raja Zahratul Azma} and Azlin Ithnin and {Abdul Razak}, {Noor Farisah} and Sardi, {Nor Hidayati} and Hussin, {Noor Hamidah}",
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T1 - Detection of α-thalassaemia in neonates on cord blood and dried blood spot samples by capillary electrophoresis

AU - Alauddin, Hafiza

AU - Langa, Mustafa

AU - Mohd Yusoff, Malisa

AU - Raja Sabudin, Raja Zahratul Azma

AU - Ithnin, Azlin

AU - Abdul Razak, Noor Farisah

AU - Sardi, Nor Hidayati

AU - Hussin, Noor Hamidah

PY - 2017/4/1

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N2 - Introduction: Haemoglobin Bart’s (Hb Bart’s) level is associated with α-thalassaemia traits in neonates, enabling early diagnosis of α-thalassaemia. The study aimed to detect and quantify the Hb Bart’s using Cord Blood (CB) and CE Neonat Fast Hb (NF) progammes on fresh and dried blood spot (DBS) specimen respectively by capillary electrophoresis (CE). Methods: Capillarys Hemoglobin (E) Kit (for CB) and Capillarys Neonat Hb Kit (for NF) were used to detect and quantify Hb Bart’s by CE in fresh cord blood and dried blood spot (DBS) specimens respectively. High performance liquid chromatography (HPLC) using the β-Thal Short Programme was also performed concurrently with CE analysis. Confirmation was obtained by multiplex ARMS Gap PCR. Results: This study was performed on 600 neonates. 32/600 (5.3%) samples showed presence of Hb Bart’s peak using the NF programme while 33/600 (5.5%) were positive with CB programme and HPLC methods. The range of Hb Bart’s using NF programme and CB programme were (0.5–4.1%) and (0.5-7.1%), respectively. Molecular analysis confirmed all positive samples possessed α-thalassaemia genetic mutations, with 23/33 cases being αα/--SEA, four -α3.7/-α3.7, two αα/-α3.7and three αα/ααCS. Fifty Hb Bart’s negative samples were randomly tested for α-genotypes, three were also found to be positive for α-globin gene mutations. Thus, resulting in sensitivity of 91.7% and 88.9% and specificity of 100% for the Capillarys Cord Blood programme and Capillarys Neonat Fast programme respectively. Conclusion: Both CE programmes using fresh or dried cord blood were useful as a screening tool for α-thalassaemia in newborns. All methods show the same specificity (100%) with variable, but acceptable sensitivities in the detection of Hb Bart.

AB - Introduction: Haemoglobin Bart’s (Hb Bart’s) level is associated with α-thalassaemia traits in neonates, enabling early diagnosis of α-thalassaemia. The study aimed to detect and quantify the Hb Bart’s using Cord Blood (CB) and CE Neonat Fast Hb (NF) progammes on fresh and dried blood spot (DBS) specimen respectively by capillary electrophoresis (CE). Methods: Capillarys Hemoglobin (E) Kit (for CB) and Capillarys Neonat Hb Kit (for NF) were used to detect and quantify Hb Bart’s by CE in fresh cord blood and dried blood spot (DBS) specimens respectively. High performance liquid chromatography (HPLC) using the β-Thal Short Programme was also performed concurrently with CE analysis. Confirmation was obtained by multiplex ARMS Gap PCR. Results: This study was performed on 600 neonates. 32/600 (5.3%) samples showed presence of Hb Bart’s peak using the NF programme while 33/600 (5.5%) were positive with CB programme and HPLC methods. The range of Hb Bart’s using NF programme and CB programme were (0.5–4.1%) and (0.5-7.1%), respectively. Molecular analysis confirmed all positive samples possessed α-thalassaemia genetic mutations, with 23/33 cases being αα/--SEA, four -α3.7/-α3.7, two αα/-α3.7and three αα/ααCS. Fifty Hb Bart’s negative samples were randomly tested for α-genotypes, three were also found to be positive for α-globin gene mutations. Thus, resulting in sensitivity of 91.7% and 88.9% and specificity of 100% for the Capillarys Cord Blood programme and Capillarys Neonat Fast programme respectively. Conclusion: Both CE programmes using fresh or dried cord blood were useful as a screening tool for α-thalassaemia in newborns. All methods show the same specificity (100%) with variable, but acceptable sensitivities in the detection of Hb Bart.

KW - Capillary electrophoresis

KW - Cord blood

KW - Neonatal α-thalassaemia screening

KW - α-thalassaemia

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