Genotyping of intron 22 inversion of factor VIII gene for diagnosis of hemophilia A by inverse-shifting polymerase chain reaction and capillary electrophoresis.

摘要: This is the first capillary electrophoresis (CE) analysis for diagnosis of hemophilia A (HA). The intron 22 inversion factor VIII gene (F8) causes 40–50 % severe bleeding disorder HA in all human populations. Consequently, identification disease-causing mutations becoming increasingly important accurate genetic counseling and prenatal diagnosis. In this study, key steps inverse-shifting polymerase chain reaction (IS-PCR) short-end injection were used more specific rapid genotyping F8. IS-PCR, three primers to amplify 512-bp amplicon wild type 584-bp patients with inversion. gel (CGE) system was performed using 1× Tris–borate–EDTA (TBE) buffer containing 0.3 % (w/v) polyethylene oxide (PEO). PCR amplicons electrokinetically injected at 10 kV 10 s a temperature 25 °C. optimal CGE applied detect F8 carriers within 5 min. Intron indeed found on some (13/35, 37.1 %). All results showed good agreement DNA sequencing method long-distance (LD-PCR). IS-PCR combined feasible efficient screening