High resolution melting analysis for a rapid identification of heterozygous and homozygous sequence changes in the MUTYH gene

Maurizio Genuardi, Rossella Tricarico, Francesca Crucianelli, Antonio Alvau, Claudio Orlando, Roberta Sestini, Francesco Tonelli, Rosa Valanzano

Risultato della ricerca: Contributo in rivistaArticolo in rivista

9 Citazioni (Scopus)

Abstract

Background: MUTYH-associated polyposis (MAP) is an autosomal recessive form of intestinal polyposis predisposing to colorectal carcinoma. High resolution melting analysis (HRMA) is a mutation scanning method that allows detection of heterozygous sequence changes with high sensitivity, whereas homozygosity for a nucleotide change may not lead to significant curve shape or melting temperature changes compared to homozygous wildtype samples. Therefore, HRMA has been mainly applied to the detection of mutations associated with autosomal dominant or X-linked disorders, while applications to autosomal recessive conditions are less common. Methods: MUTYH coding sequence and UTRs were analyzed by both HRMA and sequencing on 88 leukocyte genomic DNA samples. Twenty-six samples were also examined by SSCP. Experiments were performed both with and without mixing the test samples with wild-type DNA. Results: The results show that all MUTYH sequence variations, including G > C and A > T homozygous changes, can be reliably identified by HRMA when a condition of artificial heterozygosity is created by mixing test and reference DNA. HRMA had a sensitivity comparable to sequencing and higher than SSCP. Conclusions: The availability of a rapid and inexpensive method for the identification of MUTYH sequence variants is relevant for the diagnosis of colorectal cancer susceptibility, since the MAP phenotype is highly variable.
Lingua originaleEnglish
pagine (da-a)N/A-N/A
RivistaBMC Cancer
Volume11
DOI
Stato di pubblicazionePubblicato - 2011

Keywords

  • BASE-EXCISION-REPAIR
  • CARRIERS
  • COLORECTAL-CANCER RISK
  • GERMLINE MUTATIONS
  • LOCUS
  • MYH GENE
  • POLYPOSIS
  • SYSTEM
  • VALIDATION
  • VARIANTS

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