Detection of paternal alleles in maternal plasma for non-invasive prenatal diagnosis of beta-thalassemia: a feasibility study in southern Chinese

Eur J Obstet Gynecol Reprod Biol. 2010 May;150(1):28-33. doi: 10.1016/j.ejogrb.2010.02.016. Epub 2010 Mar 4.

Abstract

Objective: To evaluate in maternal plasma, the efficacy of detecting the paternal beta-gene mutation and informative single nucleotide polymorphisms (SNPs) linked to the paternal-mutant or -normal allele in non-invasive prenatal diagnosis (NIPND).

Study design: In 20 at-risk pregnancies, using the allele-specific arrayed primer extension (AS-APEX) technology of the previously published "Thalassemia" array, cyanine-5-deoxycytosine triphosphate (Cy5-dCTP) was incorporated into the extended strands to matched PCR-amplified maternal plasma DNA templates, to detect both the paternal beta-gene mutation and informative paternal SNPs.

Results: Sensitivity experiment showed that 5pg DNA as starting template gave detectable signals on the array. In 13 cases (65%), the paternal-derived beta-gene mutation and/or informative mutant-associated SNP were detected. A subsequent invasive procedure was required to determine if the fetus had a beta-thalassemia (thal) major or minor genotype. In 3 cases (15%), absence of the paternal mutant or mutant-associated SNP excluded a beta-thal major fetus; while in 4 cases (20%), this approach was non-discriminative as both parents carry the same mutation without any informative SNP.

Conclusion: This approach was useful in 16 out of 20 (80%) pregnancies at risk for beta-thal in southern Chinese.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alleles*
  • Asian People / genetics
  • Carbocyanines
  • Deoxycytosine Nucleotides
  • Fathers
  • Feasibility Studies
  • Female
  • Humans
  • Male
  • Polymerase Chain Reaction / methods
  • Polymorphism, Single Nucleotide
  • Pregnancy
  • Prenatal Diagnosis / methods*
  • beta-Globins / genetics
  • beta-Thalassemia / diagnosis*
  • beta-Thalassemia / genetics

Substances

  • Carbocyanines
  • Cy5-dCTP
  • Deoxycytosine Nucleotides
  • beta-Globins