Detection of heterozygous large deletions in the antithrombin gene using multiplex polymerase chain reaction and denatured high performance liquid chromatography.
The present study reports a method for the easy, rapid and cost effective detection of heterozygous large deletions. As a model gene all exons of the antithrombin gene were amplified in a one tube multiplex polymerase chain reaction (PCR) and the products separated according to their size by reverse-phase ion-pair high performance liquid chromatography. A significant reduction in the height of a peak in the probandOs sample compared to in the control indicates the presence of a large deletion of the corresponding allele. Using this approach we identified heterozygous deletions in four patients: the deletions affected exons 1 and 2, exon 7 and the whole antithrombin gene.
Pavlova
Anna
A
Institute of Transfusion Medicine and Immunohaematolgy, DRK Blood Donor Service Baden-Württemberg-Hessen, 60526 Frankfurt/M, Germany.
El-Maarri
Osman
O
Luxembourg
Beate
B
Lindhoff-Last
Edelgard
E
Kochhan
Lothar
L
Bruhn
Hans-Dietrich
HD
Delev
Daniel
D
Watzka
Matthias
M
Seifried
Erhard
E
Oldenburg
Johannes
J
D000990:Antithrombin III / Q000235:genetics*; D002851:Chromatography, High Pressure Liquid; D005091:Exons; D017353:Gene Deletion; D006580:Genetic Carrier Screening / Q000379:methods*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D017384:Sequence Deletion
D000990:Antithrombin III