skip to main content

Comparison of multiple genotyping methods for the identification of the cancer predisposing founder mutation p.R337H in TP53

Fitarelli-Kiehl, Mariana ; Macedo, Gabriel S ; Schlatter, Rosane Paixão ; Koehler-Santos, Patricia ; Matte, Ursula Da Silveira ; Ashton-Prolla, Patricia ; Giacomazzi, Juliana

Genetics and Molecular Biology, 01 June 2016, Vol.39(2), pp.203-209 [Periódico revisado por pares]

Texto completo disponível

Citações Citado por
  • Título:
    Comparison of multiple genotyping methods for the identification of the cancer predisposing founder mutation p.R337H in TP53
  • Autor: Fitarelli-Kiehl, Mariana ; Macedo, Gabriel S ; Schlatter, Rosane Paixão ; Koehler-Santos, Patricia ; Matte, Ursula Da Silveira ; Ashton-Prolla, Patricia ; Giacomazzi, Juliana
  • Assuntos: Biochemistry & Molecular Biology ; Genetics & Heredity ; Tp53-P.R337h ; Rflp ; Taqman ; Hrm ; Sanger Sequencing ; Biology
  • É parte de: Genetics and Molecular Biology, 01 June 2016, Vol.39(2), pp.203-209
  • Descrição: Abstract Germline mutations in the TP53 gene are associated with Li-Fraumeni and Li-Fraumeni-Like Syndromes, characterized by increased predisposition to early-onset cancers. In Brazil, the prevalence of the TP53-p.R337H germline mutation is exceedingly high in the general population and in cancer-affected patients, probably as result of a founder effect. Several genotyping methods are used for the molecular diagnosis of LFS/LFL, however Sanger sequencing is still considered the gold standard. We compared performance, cost and turnaround time of Sanger sequencing, PCR-RFLP, TaqMan-PCR and HRM in the p.R337H genotyping. The performance was determined by analysis of 95 genomic DNA samples and results were 100% concordant for all methods. Sequencing was the most expensive method followed by TaqMan-PCR, PCR-RFLP and HRM. The overall cost of HRM increased with the prevalence of positive samples, since confirmatory sequencing must be performed when a sample shows an abnormal melting profile, but remained lower than all other methods when the mutation prevalence was less than 2.5%. Sequencing had the highest throughput and the longest turnaround time, while TaqMan-PCR showed the lowest turnaround and hands-on times. All methodologies studied are suitable for the detection of p.R337H and the choice will depend on the application and clinical scenario.
  • Idioma: Português

Buscando em bases de dados remotas. Favor aguardar.