title: Rapid detection of α-thalassaemia variants using droplet digital PCR.
abstract: BACKGROUND
  Alpha thalassaemia is a highly prevalent disease globally and is a well-known public health problem in Malaysia. The deletional forms of the mutation are the most common forms found in alpha thalassaemia. The three most common deletional alpha thalassaemia found in this region include --(SEA) deletion, -α(3.7) rightward and -α(4.2) leftward deletions. The prevalence rate of triplication alpha cases such as ααα(anti3.7) and ααα(anti4.2) is not known in Malaysia although it plays a role in exacerbating the clinical phenotypes in beta thalassaemia carriers. Recently, there have been more reported cases of rare alpha thalassaemia mutations due to the advancement of molecular techniques involved in thalassaemia detections. Therefore, it is essential to develop a new method which allows the detection of different alpha thalassaemia mutations including the rare ones simultaneously and accurately.
  
  
  METHODS
  The purpose of this study was to design an assay for the detection of triplications, common and rare deletional alpha thalassaemia using droplet digital PCR (ddPCR).
  
  
  RESULTS
  This is a quantitative detection method to measure the changes of copy number which can detect deletions, duplications and triplications of the alpha globin gene simultaneously.
  
  
  CONCLUSIONS
  In conclusion, ddPCR is an alternative method for rapid detection of alpha thalassaemia variants in Malaysia.
authors.affiliation: Department of Pathology, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Malaysia.
authors.forename: T-Y
authors.initials: TY
authors.lastname: Lee
authors.affiliation: Department of Pathology, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Malaysia.
authors.forename: M-I
authors.initials: MI
authors.lastname: Lai
authors.affiliation: Malaysian Research Institute on Ageing, Universiti Putra Malaysia, Serdang, Malaysia.
authors.forename: V
authors.initials: V
authors.lastname: Ramachandran
authors.affiliation: Department of Biomedical Science, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia.
authors.forename: J A M A
authors.initials: JA
authors.lastname: Tan
authors.affiliation: Department of Biomedical Science, Faculty of Science, Universiti Tunku Abdul Rahman, Kampar, Malaysia.
authors.forename: L-K
authors.initials: LK
authors.lastname: Teh
authors.affiliation: Department of Pathology, Hospital Kuala Lumpur, Kuala Lumpur, Malaysia.
authors.forename: R
authors.initials: R
authors.lastname: Othman
authors.affiliation: Department of Pathology, Hospital Kuala Lumpur, Kuala Lumpur, Malaysia.
authors.forename: N H
authors.initials: NH
authors.lastname: Hussein
authors.affiliation: Department of Pathology, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Malaysia.
authors.forename: E
authors.initials: E
authors.lastname: George
mesh_terms: D056915:DNA Copy Number Variations; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D008296:Malaysia; D017720:Molecular Epidemiology; D009154:Mutation; D010641:Phenotype; D016133:Polymerase Chain Reaction / Q000379:methods*; D015995:Prevalence; D017384:Sequence Deletion; D017085:alpha-Thalassemia / Q000453:epidemiology / Q000235:genetics*
keywords: Malaysia; alpha thalassaemia; ddPCR; detection; triplication