Quantitative polymerase chain reaction for the rapid prenatal diagnosis of homozygous alpha-thalassaemia (Hb Barts hydrops fetalis)

Br J Haematol. 2001 Nov;115(2):341-6. doi: 10.1046/j.1365-2141.2001.03112.x.

Abstract

A quantitative polymerase chain reaction (Q-PCR) method based on the TaqMan technology has been devised for the prenatal diagnosis of homozygous alpha*-thalassaemia (south-east Asian type deletion). Primers and TaqMan probes were designed to specifically amplify an alpha*-thal chromosomal fragment or a normal alpha-chromosomal fragment. Variations in input target DNA in individual sample wells were normalized by the simultaneous amplification of a beta-actin gene fragment and results expressed as a ratio to that of beta-actin. There was no overlap of the data between the homozygous alpha*-thal, alpha*-thal and normal subjects. Up to 5% maternal DNA (alpha*-thal) contamination did not affect the specificity of the result. In 31 prenatal diagnoses, the result using Q-PCR compared favourably with the gold standard of Southern hybridization of alpha-genes.

Publication types

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

MeSH terms

  • Actins / genetics
  • Female
  • Homozygote
  • Humans
  • Hydrops Fetalis / diagnosis*
  • Hydrops Fetalis / genetics
  • Polymerase Chain Reaction / methods
  • Pregnancy
  • Prenatal Diagnosis / methods*
  • alpha-Thalassemia / diagnosis*
  • alpha-Thalassemia / genetics

Substances

  • Actins