EMMA, a cost- and time-effective diagnostic method for simultaneous detection of point mutations and large-scale genomic rearrangements: application to BRCA1 and BRCA2 in 1,525 patients.
The detection of unknown mutations remains a serious challenge and, despite the expected benefits for the patient's health, a large number of genes are not screened on a routine basis. We present the diagnostic application of EMMA (Enhanced Mismatch Mutation Analysis(®) , Fluigent, Paris, France), a novel method based on heteroduplex analysis by capillary electrophoresis using innovative matrices. BRCA1 and BRCA2 were screened for point mutations and large rearrangements in 1,525 unrelated patients (372 for the validation step and 1,153 in routine diagnosis) using a single analytical condition. Seven working days were needed for complete BRCA1/2 screening in 30 patients by one technician (excluding DNA extraction and sequencing). A total of 137 mutations were found, including a BRCA2 duplication of exons 19 and 20, previously missed by Comprehensive BRACAnalysis(®) . The mutation detection rate was 11.9%, which is consistent with patient inclusions. This study therefore suggests that EMMA represents a valuable short-term and midterm option for many diagnostic laboratories looking for an easy, reliable, and affordable strategy, enabling fast and sensitive analysis for a large number of genes.
Service de Génétique Oncologique, Institut Curie, Paris, France.
Virginie
V
Caux-Moncoutier
Laurent
L
Castéra
Carole
C
Tirapo
Dorothée
D
Michaux
Marie-Alice
MA
Rémon
Anthony
A
Laugé
Etienne
E
Rouleau
Antoine
A
De Pauw
Bruno
B
Buecher
Marion
M
Gauthier-Villars
Jean-Louis
JL
Viovy
Dominique
D
Stoppa-Lyonnet
Claude
C
Houdayer
D019313:BRCA1 Protein / Q000032:analysis / Q000235:genetics*; D024682:BRCA2 Protein / Q000032:analysis / Q000235:genetics*; D001943:Breast Neoplasms / Q000235:genetics; D002869:Chromosome Aberrations; D003362:Cost-Benefit Analysis; D004252:DNA Mutational Analysis / Q000191:economics / Q000379:methods*; D004274:DNA, Recombinant; D019075:Electrophoresis, Capillary; D005260:Female; D016368:Frameshift Mutation; D019398:Genes, BRCA1; D024522:Genes, BRCA2; D005820:Genetic Testing / Q000379:methods*; D057166:High-Throughput Screening Assays / Q000379:methods; D006801:Humans; D020125:Mutation, Missense; D010051:Ovarian Neoplasms / Q000235:genetics; D017354:Point Mutation
D019313:BRCA1 Protein; C492913:BRCA1 protein, human; D024682:BRCA2 Protein; C551750:BRCA2 protein, human; D004274:DNA, Recombinant