title: Quantitative PCR analysis reveals a high incidence of large intragenic deletions in the FANCA gene in Spanish Fanconi anemia patients.
abstract: Fanconi anaemia is an autosomal recessive disease characterized by chromosome fragility, multiple congenital abnormalities, progressive bone marrow failure and a high predisposition to develop malignancies. Most of the Fanconi anaemia patients belong to complementation group FA-A due to mutations in the FANCA gene. This gene contains 43 exons along a 4.3-kb coding sequence with a very heterogeneous mutational spectrum that makes the mutation screening of FANCA a difficult task. In addition, as the FANCA gene is rich in Alu sequences, it was reported that Alu-mediated recombination led to large intragenic deletions that cannot be detected in heterozygous state by conventional PCR, SSCP analysis, or DNA sequencing. To overcome this problem, a method based on quantitative fluorescent multiplex PCR was proposed to detect intragenic deletions in FANCA involving the most frequently deleted exons (exons 5, 11, 17, 21 and 31). Here we apply the proposed method to detect intragenic deletions in 25 Spanish FA-A patients previously assigned to complementation group FA-A by FANCA cDNA retroviral transduction. A total of eight heterozygous deletions involving from one to more than 26 exons were detected. Thus, one third of the patients carried a large intragenic deletion that would have not been detected by conventional methods. These results are in agreement with previously published data and indicate that large intragenic deletions are one of the most frequent mutations leading to Fanconi anaemia. Consequently, this technology should be applied in future studies on FANCA to improve the mutation detection rate.
authors.affiliation: Universitat Autònoma de Barcelona, Barcelona, Spain.
authors.forename: E
authors.initials: E
authors.lastname: Callén
authors.forename: M D
authors.initials: MD
authors.lastname: Tischkowitz
authors.forename: A
authors.initials: A
authors.lastname: Creus
authors.forename: R
authors.initials: R
authors.lastname: Marcos
authors.forename: J A
authors.initials: JA
authors.lastname: Bueren
authors.forename: J A
authors.initials: JA
authors.lastname: Casado
authors.forename: C G
authors.initials: CG
authors.lastname: Mathew
authors.forename: J
authors.initials: J
authors.lastname: Surrallés
mesh_terms: D020087:Alu Elements; D004252:DNA Mutational Analysis; D004268:DNA-Binding Proteins / Q000235:genetics*; D005091:Exons / Q000235:genetics; D005199:Fanconi Anemia / Q000453:epidemiology / Q000235:genetics*; D052217:Fanconi Anemia Complementation Group A Protein; D017353:Gene Deletion; D018740:Genetic Heterogeneity; D006579:Heterozygote; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D013030:Spain / Q000453:epidemiology
chemical_list: D004268:DNA-Binding Proteins; C498471:FANCA protein, human; D052217:Fanconi Anemia Complementation Group A Protein