title: Analysis of β/α globin ratio by using relative qRT-PCR for diagnosis of beta-thalassemia carriers.
abstract: BACKGROUND
  Current routine tests for premarital screening of β-thalassemia carriers are not applicable for diagnosis of rare atypical minor β-thalassemia cases. A more specialized laboratory evaluation for them is the measurement of β/α chain synthesis ratio with the assistance of radioactive amino acids. This method is also no longer routinely accessible. Consequently it is required to establish a rapid, trouble-free, and reliable method that encompasses all the cases of β-thalassemia carriers. Therefore we have determined β/α-globin mRNA ratio by applying relative qRT-PCR in various β-thalassemia patients.
  
  
  METHODS
  Reticulocytes RNA extraction and subsequent cDNA synthesis were performed, followed by relative qRT-PCR for α- and β-globin chain genes and β-actin gene as an endogenous reference. β/α-Globin gene ratio was then evaluated with the Pfaffl method.
  
  
  RESULTS
  The mean of β/α ratio was 0.99, 0.81, 0.69, and 0.69 for normal population, minor, intermediate, and major β-thalassemia, respectively. Approximately 6% of cases with minor thalassemia RBC index and normal HbA2 and having a decreased β/α ratio were located in the minor β-thalassemia group. The mean of β/α mRNA ratio in normal individuals and minor β-thalassemia was significantly different with all other groups (P-value < 0.05). Nevertheless, there was no such association between β/α mRNA ratio in major and intermediate β-thalassemia.
  
  
  CONCLUSIONS
  According to the significant differences achieved, no overlapping between minor β-thalassemia and normal group, capability of diagnosing atypical minor β-thalassemia, and accessibility of this technique, we can declare that this method could be suggested as a routine premarital screening test for β-thalassemia carriers.
authors.affiliation: Diagnostic Laboratory Sciences and Technology Research Center, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran.
authors.forename: Reza
authors.initials: R
authors.lastname: Ranjbaran
authors.forename: Mohammad Ali
authors.initials: MA
authors.lastname: Okhovat
authors.forename: Arash
authors.initials: A
authors.lastname: Mobarhanfard
authors.forename: Farzaneh
authors.initials: F
authors.lastname: Aboualizadeh
authors.forename: Mozhdeh
authors.initials: M
authors.lastname: Abbasi
authors.forename: Leili
authors.initials: L
authors.lastname: Moezzi
authors.forename: Habib Allah
authors.initials: HA
authors.lastname: Golafshan
authors.forename: Abbas
authors.initials: A
authors.lastname: Behzad-Behbahani
authors.forename: Mandana
authors.initials: M
authors.lastname: Bagheri
authors.forename: Sedigheh
authors.initials: S
authors.lastname: Sharifzadeh
mesh_terms: D000293:Adolescent; D002648:Child; D002675:Child, Preschool; D006580:Genetic Carrier Screening / Q000379:methods*; D005820:Genetic Testing / Q000379:methods; D006801:Humans; D007223:Infant; D011291:Premarital Examinations / Q000379:methods; D012333:RNA, Messenger / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D055542:alpha-Globins / Q000235:genetics*; D055544:beta-Globins / Q000235:genetics*; D017086:beta-Thalassemia / Q000175:diagnosis* / Q000235:genetics
chemical_list: D012333:RNA, Messenger; D055542:alpha-Globins; D055544:beta-Globins
keywords: premarital screening; relative qRT-PCR; thalassemia