Comparison of multiple genotyping methods for the identification of the cancer predisposing founder mutation p.R337H in TP53
dc.contributor.author | Kiehl, Mariana Fitarelli | pt_BR |
dc.contributor.author | Macedo, Gabriel de Souza | pt_BR |
dc.contributor.author | Schlatter, Rosane Paixão | pt_BR |
dc.contributor.author | Santos, Patrícia Koehler dos | pt_BR |
dc.contributor.author | Matte, Ursula da Silveira | pt_BR |
dc.contributor.author | Prolla, Patrícia Ashton | pt_BR |
dc.contributor.author | Giacomazzi, Juliana | pt_BR |
dc.date.accessioned | 2016-08-20T02:14:58Z | pt_BR |
dc.date.issued | 2016 | pt_BR |
dc.identifier.issn | 1415-4757 | pt_BR |
dc.identifier.uri | http://hdl.handle.net/10183/147480 | pt_BR |
dc.description.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. | en |
dc.format.mimetype | application/pdf | pt_BR |
dc.language.iso | eng | pt_BR |
dc.relation.ispartof | Genetics and molecular biology. Ribeirão Preto, SP. Vol. 39, no. 2 (Jun. 2016), p. 203-209 | pt_BR |
dc.rights | Open Access | en |
dc.subject | TP53-p.R337H | en |
dc.subject | Neoplasias | pt_BR |
dc.subject | Predisposição genética para doença | pt_BR |
dc.subject | RFLP | en |
dc.subject | TaqMan | en |
dc.subject | HRM | en |
dc.subject | Sanger sequencing | en |
dc.title | Comparison of multiple genotyping methods for the identification of the cancer predisposing founder mutation p.R337H in TP53 | pt_BR |
dc.type | Artigo de periódico | pt_BR |
dc.identifier.nrb | 000997097 | pt_BR |
dc.type.origin | Nacional | pt_BR |
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