Molecular Prenatal Diagnosis of -Thalassemia Using Real-Time and Multiplex Polymerase Chain Reaction Methods

Molecular analysis of two fetuses at high risk of -thalassemia (-thal), and their family members, was performed using real-time polymerase chain reaction (PCR) with SYBR Green 1 (SYBR-PCR) dye combined with dissociation curve analysis and multiplex PCR (m-PCR) and DNA sequencing techniques. The genotype of the fetus from one family was - -SEA/- -SEA (Southeast Asian deletion), which produces hydrops fetalis syndrome. The genotype of the parents was - -SEA/. A boy with Hb H disease and his sibling fetus from the other family had the genotype - -SEA/CS [the Hb Constant Spring (CS) mutation: 142, TermGln (TAACAA in 2)] and / (normal), respectively. The diagnosis, based on SYBR-PCR combined with dissociation curve analysis, was in agreement with the results from the m-PCR method. This indicates that these are alternative and reliable assays for the molecular diagnosis of deletional -thal.