Quantitative polmerase chain reaction for the rapid prenatal diagnosis of homozygous α‐thalassaemia (Hb Barts hydrops fetalis)

• Published in: British journal of haematology Vol. 115; no. 2; pp. 341 - 346
• Main Authors: Chan, Vivian, Yip, Ben, Lam, Y. H., Tse, H. Y., Wong, H. S., Chan, T. K.
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Science Ltd 01.11.2001; Blackwell
• Subjects: Anemias. Hemoglobinopathies; Biological and medical sciences; Diseases of red blood cells; Hematologic and hematopoietic diseases; hydrops fetalis; Medical sciences; prenatal diagnosis; Q‐PCR; α°‐thalassaemia; Human; Hemoglobinopathy; Pregnancy disorders; Hemopathy; Genetic disease; Prenatal; Polymerase chain reaction; Fetal diseases; Hemolytic anemia; Hydrops fetalis; α-Thalassemia; Diagnosis; Technique; Molecular biology; Quantitative analysis
• ISSN: 0007-1048; 1365-2141
• DOI: 10.1046/j.1365-2141.2001.03112.x

A quantitative polymerase chain reaction (Q‐PCR) method based on the TaqMan technology has been devised for the prenatal diagnosis of homozygous α°−thalassaemia (south‐east Asian type deletion). Primers and TaqMan probes were designed to specifically amplify an α°‐thal chromosomal fragment or a normal α‐chromosomal fragment. Variations in input target DNA in individual sample wells were normalized by the simultaneous amplification of a β‐actin gene fragment and results expressed as a ratio to that of β‐actin. There was no overlap of the data between the homozygous α°‐thal, α°‐thal and normal subjects. Up to 5% maternal DNA (α°‐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 α‐genes.