Single-strand conformation polymorphism (SSCP) analysis of the molecular pathology of hemophilia B

Hum Mutat. 1993;2(5):355-61. doi: 10.1002/humu.1380020506.


In the present study, we report the application of polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) analysis to the screening of seven functionally important factor IX gene (FIX) regions (total length 2.66 kb) in 9 unrelated haemophilia B patients of Portuguese or African origin. In eight of the patients an altered migration pattern of single-stranded DNA was observed. Direct sequencing of the relevant DNA fragments unveiled the following sequence alterations: two novel mutations, namely FIXBarcelos Thr-380-Pro and FIXLousada 9bp insertion at position 31,309 or 31,318; five mutations previously reported in other ethnic groups (FIXPorto Arg-145-His, FIXLuanda Gly-207-Arg, FIXPenafiel Arg-248-Gln, FIXSesimbra Arg-333-Gln, FIXCascais Arg-333-Stop); and a normal variant, G-->T transvertion at position 6,596 in intron 2. We propose hypothetical models for the generation of the 9 bp duplication (FIXLousada). We have performed molecular modeling studies in order to predict the structure of the variant FIX molecules.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • DNA / genetics*
  • DNA, Single-Stranded / genetics
  • Factor IX / genetics*
  • Hemophilia B / genetics*
  • Humans
  • Molecular Sequence Data
  • Mutation / genetics
  • Nucleic Acid Conformation
  • Polymerase Chain Reaction / methods
  • Polymorphism, Genetic / genetics*


  • DNA, Single-Stranded
  • Factor IX
  • DNA