Rapid Detection of Fetal Mendelian Disorders: Thalassemia and Sickle Cell Syndromes

Methods Mol Biol. 2019;1885:207-219. doi: 10.1007/978-1-4939-8889-1_14.


The inherited disorders of hemoglobin synthesis constitute the most common monogenic diseases worldwide. The clinical severity of β-thalassemia major and the sickle cell syndromes targets them as priority genetic diseases for prevention programs, which incorporates population screening to identify heterozygotes, with the option of prenatal diagnosis for carrier couples. Rapid genotype characterization is fundamental in the diagnostic laboratory, especially when offering prenatal diagnosis. The application of real-time PCR provides a means for rapid and potentially high throughput assays, without compromising accuracy. It has several advantages over end-point PCR analysis, including the elimination of post-PCR processing steps and a wide dynamic range of detection with a high degree of sensitivity. Although there are over 200 mutations associated with the β-thalassemia and sickle cell syndromes, the relatively small size of the β-, HBB gene (less than 2000 base-pairs) and the close proximity of most mutations facilitates the design of a minimal number of real-time PCR assays using the LightCycler™ system, which are capable of detecting the majority of most common β-gene mutations world-wide. These assays are highly appropriate for rapid genotyping of parental and fetal DNA samples with respect to β-thalassemia and sickle cell syndromes.

Keywords: Prenatal diagnosis; Real-time PCR; β-thalassemia and sickle cell syndromes.

MeSH terms

  • Amniotic Fluid / cytology
  • Anemia, Sickle Cell / diagnosis*
  • Anemia, Sickle Cell / genetics*
  • Chorionic Villi Sampling
  • DNA Mutational Analysis
  • Genetic Association Studies
  • Genetic Predisposition to Disease
  • Genetic Testing* / methods
  • Humans
  • Mutation
  • Prenatal Diagnosis* / methods
  • Real-Time Polymerase Chain Reaction
  • beta-Globins / genetics
  • beta-Thalassemia / diagnosis*
  • beta-Thalassemia / genetics*


  • beta-Globins