Direct carrier detection in hemophilia B kindreds: Use of modified primers (mutagenic primers) for enzymatic amplification of the factor IX gene

Tadashi Matsushita, Mitsune Tanimoto, Koji Yamamoto, Isamu Sugiura, Junki Takamatsu, Tadashi Kamiya, Hidehiko Saito

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Rapid direct detection of point mutations in hemophilia B kindreds was performed by analyzing the restriction fragments of the factor IX gene amplified by polymerase chain reaction (PCR). The family members of the two patients, HB 5 and HB 6, whose mutant factor IX gene had previously been defined by sequence analysis, were examined in this study. Since there are no restriction endonucleases available for detecting each mutation directly, we designed modified primers which were substituted one nucleotide near the mutated positions. Following PCR with these primers, new Mbo I or Alu I cleavage sites for normal alleles were created. In each family, the restriction fragment revealed the carriership of each patient's mother. This simple methodology to detect mutations of interest is useful for genetic counseling in sporadic cases of hemophilia B.

Original languageEnglish
Pages (from-to)355-361
Number of pages7
JournalThrombosis Research
Volume63
Issue number3
DOIs
Publication statusPublished - Aug 1 1991

Keywords

  • Carrier
  • Factor IX
  • Hemophilia B
  • Polymerase chain reaction
  • Prenatal diagnosis

ASJC Scopus subject areas

  • Hematology

Fingerprint Dive into the research topics of 'Direct carrier detection in hemophilia B kindreds: Use of modified primers (mutagenic primers) for enzymatic amplification of the factor IX gene'. Together they form a unique fingerprint.

Cite this