title: Molecular characterization of hemoglobin D Punjab traits and clinical-hematological profile of the patients.
abstract: OBJECTIVE
  Hemoglobin (Hb) D hemoglobinopathies are widespread diseases in northwestern India and usually present with mild hemolytic anemia and mild to moderate splenomegaly. The heterozygous form of Hb D is clinically silent, but coinheritance of Hb D with Hb S or beta-thalassemia produces clinically significant conditions like thalassemia intermedia of moderate severity. Under heterozygous conditions with coinheritance of alpha and beta-thalassemia, patients show a degree of clinical variability. Thus, our aim was to molecularly characterize the Hb D trait among individuals who were clinically symptomatic because of co-inheritance of alpha deletions or any beta-globin gene mutations.
  
  
  METHODS
  This was a cross-sectional study conducted in an autonomous tertiary-care hospital.
  
  
  METHODS
  Complete blood count and red cell indices were measured using an automated cell analyzer. Quantitative assessment of hemoglobin Hb F, Hb A, Hb A2 and Hb D was performed by means of high performance liquid chromatography (HPLC). DNA extraction was done using the phenol-chloroform method. Molecular analyses on common alpha deletions and common beta mutations were done using the Gap polymerase chain reaction and Amplification Refractory Mutation System, respectively.
  
  
  RESULTS
  We evaluated 30 patients and found clinical variation in the behavior of Hb D traits. In six patients, the Hb D traits were clinically symptomatic and behaved like those of thalassemia intermedia. Molecular characterization showed that three out of these six were IVS-1-5 positive.
  
  
  CONCLUSIONS
  HPLC may not be the gold standard for diagnosing symptomatic Hb D Punjab traits. Hence, standard confirmation should include molecular studies.
authors.affiliation: Department of Hematology, All India Institute of Medical Sciences, New Delhi, India.
authors.forename: Sanjay
authors.initials: S
authors.lastname: Pandey
authors.forename: Rahasya Mani
authors.initials: RM
authors.lastname: Mishra
authors.forename: Sweta
authors.initials: S
authors.lastname: Pandey
authors.forename: Vineet
authors.initials: V
authors.lastname: Shah
authors.forename: Renu
authors.initials: R
authors.lastname: Saxena
mesh_terms: D000293:Adolescent; D000328:Adult; D002648:Child; D002675:Child, Preschool; D002853:Chromatography, Liquid; D003430:Cross-Sectional Studies; D005260:Female; D006453:Hemoglobinopathies / Q000097:blood / Q000235:genetics*; D006454:Hemoglobins / Q000032:analysis / Q000235:genetics*; D006455:Hemoglobins, Abnormal / Q000032:analysis / Q000235:genetics*; D006801:Humans; D007194:India; D007223:Infant; D008297:Male; D009154:Mutation / Q000235:genetics; D016133:Polymerase Chain Reaction; D055815:Young Adult
chemical_list: D006454:Hemoglobins; D006455:Hemoglobins, Abnormal; C026237:hemoglobin D Punjab