Breast Cancer Research and Treatment

, Volume 115, Issue 2, pp 405–414 | Cite as

High-resolution melting analysis for rapid screening of BRCA1 and BRCA2 Spanish mutations

  • Inmaculada de Juan
  • Eva Esteban
  • Sarai Palanca
  • Eva Barragán
  • Pascual BoluferEmail author


The majority of BRCA1 and BRCA2 mutation detection procedures include screening methods, all of which are time-consuming. High-resolution melting (HRM) is a promising pre-screening method of gene scanning that combines simplicity and rapid identification of genetic variants. We evaluated HRM in the screening of BRCA1/2 Spanish mutations. We studied 40 BRCA1 and 47 BRCA2 DNA samples with different Spanish mutations. We included a group of 20 unknown DNAs from patients with sporadic breast cancer (BC). The assay was performed with the LightCycler® 480 Instrument (Roche). The HRM discriminates all the BRCA1/2 Spanish mutations studied from wild-type DNA. Besides, 54 out of 87 mutations were clearly differentiated from each other. In sporadic BC 11 polymorphisms and three unclassified variants were found in both genes. HRM is a valuable method for rapid screening of BRCA1/2 Spanish mutations and is capable of differentiating new genetic variants in PCR products.


Hereditary breast and ovarian cancer BRCA1 BRCA2 Screening Mutations High-resolution melting 



This work has been founded by the grants FIS PI060505 (Instituto de Salud Carlos III) and AP-042/07 (Consellería de Sanitat). We also express our gratitude to the “Fundación para Investigación La Fe” for having granted Inmaculada de Juan (Bch Sc and Specialist in Clinical Chemistry) in a research project on sporadic breast cancer, which made possible her participation in the present study. We should thank and name the colleagues who kindly provided DNA samples containing mutations, and those others that send us patient’s samples for the present: Cristina Miner (Laboratorio de Genética del Cáncer, Instituto de Biología y Genética Molecular, Valladolid), Orlan Diez (Servicio de Genética, Hospital de la Santa Creu i San Pau, Barcelona), Trinidad Caldés (Laboratorio de Biología Molecular, Hospital Clínico San Carlos, Madrid), Rogelio González (Centro de Investigación del Cáncer, Salamanca), Javier Benítez (Centro Nacional de Investigaciones Oncológicas, Madrid), Ana Vega (Unidad de Medicina Molecular, Fundación Pública Galega de Medicina Xenómica-SERGAS, Hospital Clínico Universitario de Santiago de Compostela, Santiago de Compostela), Ismael Aznar (Servicio de Cirugía del Hospital La Fe), Carlos Vázquez (Servicio de Cirugía del IVO), Isabel Chirivella (Unidad de Consejo Genético en Cáncer Hospital Clínico Universitario, Valencia), Ángel Segura (Unidad de Consejo Genético en Cáncer Hospital Universitario La Fe, Valencia), Carmen Guillén (Unidad de Consejo Genético en Cáncer Hospital General de Elche, Alicante), Eduardo Martínez (Unidad de Consejo Genético en Cáncer Consorcio Hospital Provincial, Castellón), Dolores Cuevas (Servicio de Protocolización e Integración Asistencial, Dirección General de Asistencia Sanitaria, Agencia Valenciana de Salud), Dolores Salas (Oficina del Plan de Cáncer, Dirección General de Salud Pública, Consellería de Sanitat de Comunidad Valenciana).

Supplementary material

10549_2008_73_MOESM1_ESM.pdf (132 kb)
Difference plots of BRCA1 Spanish mutations. Ex. = exon. For PCRNumber and conditions see Table 1 (PDF 133 kb)
10549_2008_73_MOESM2_ESM.pdf (161 kb)
Difference plots of BRCA 2 Spanish mutations. Ex. = exon. For PCRNumber and conditions see Table 2 (PDF 162 kb)


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Copyright information

© Springer Science+Business Media, LLC. 2008

Authors and Affiliations

  • Inmaculada de Juan
    • 1
  • Eva Esteban
    • 1
  • Sarai Palanca
    • 1
  • Eva Barragán
    • 1
  • Pascual Bolufer
    • 1
    Email author
  1. 1.Laboratory of Molecular BiologyUniversity Hospital La FeValenciaSpain

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