| Registro completo |
| Provedor de dados: |
Genet. Mol. Biol.
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| País: |
Brazil
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| Título: |
Comparison of multiple genotyping methods for the identification of the cancer predisposing founder mutation p.R337H in TP53
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| Autores: |
Fitarelli-Kiehl,Mariana
Macedo,Gabriel S.
Schlatter,Rosane Paixão
Koehler-Santos,Patricia
Matte,Ursula da Silveira
Ashton-Prolla,Patricia
Giacomazzi,Juliana
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| Data: |
2016-06-01
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| Ano: |
2016
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| Palavras-chave: |
TP53-p.R337H
RFLP
TaqMan
HRM
Sanger Sequencing
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| Resumo: |
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.
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| Tipo: |
Info:eu-repo/semantics/article
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| Idioma: |
Inglês
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| Identificador: |
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572016000200203
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| Editor: |
Sociedade Brasileira de Genética
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| Relação: |
10.1590/1678-4685-gmb-2014-0351
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| Formato: |
text/html
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| Fonte: |
Genetics and Molecular Biology v.39 n.2 2016
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| Direitos: |
info:eu-repo/semantics/openAccess
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