Analysis of real-time PCR cycle threshold of alpha-thalassemia-1 Southeast Asian type deletion using fetal cell-free DNA in maternal plasma for noninvasive prenatal diagnosis of Bart's hydrops fetalis

J Med Assoc Thai. 2010 Nov;93(11):1243-8.

Abstract

Background: Noninvasive prenatal diagnosis based on detection of fetal cell-free DNA is hampered when mother and father are both carriers for the same autosomal recessive mutation.

Objective: To compare the diagnosis of Bart's hydrops fetalis using conventional Gap-PCR analysis of fetal cells/tissues with the measurement of quantitative difference (deltaCp) between alpha-thalassemia-1 SEA type deletion gene (C(T-mutant)) and wild type alpha-globin gene (C(T-wild type)) in plasma of pregnancies by using the Taqman real-time quantitative PCR.

Material and method: Plasma DNA samples were collected from three groups of pregnancies whose fetuses have known thalasemia status (7 normal, 11 heterozygote alpha-thalassemia-1 SEA type deletion, and 7 Bart's hydrops fetalis). The alpha-thalassemia-1 SEA type deletion gene and wild type alpha-globin gene were quantified by using Taqman real-time quantitative PCR and then the delta C(T) was analyzed by subtracting the C(T-mutant) from C(T-wild type).

Results: Mean deltaC(T) values were not significantly different among the three groups. However, women whose fetuses were diagnosed as Bart's hydrops fetalis had a higher proportion (43%) of plasma DNA samples that had negative deltaC(T) value than women whose fetuses were diagnosed as normal or heterozygote alpha-thalassemia-1 SEA type deletion (0 and 27%, respectively).

Conclusion: Further investigations are needed to improve the diagnosis of Bart's hydrops fetalis using fetal cell-free DNA.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alpha-Globulins / genetics*
  • Asian People / genetics
  • DNA / blood
  • DNA / isolation & purification
  • Female
  • Fetus*
  • Hemoglobins, Abnormal / genetics
  • Humans
  • Hydrops Fetalis / diagnosis*
  • Hydrops Fetalis / genetics
  • Mothers
  • Plasma / chemistry
  • Polymerase Chain Reaction / methods*
  • Pregnancy
  • Pregnancy Trimester, First
  • Pregnancy Trimester, Second
  • Prenatal Diagnosis / methods*
  • Sequence Deletion
  • alpha-Thalassemia / blood
  • alpha-Thalassemia / diagnosis*
  • alpha-Thalassemia / genetics

Substances

  • Alpha-Globulins
  • Hemoglobins, Abnormal
  • DNA
  • hemoglobin Bart's