TY - JOUR
T1 - A strategy for molecular diagnostics of Fanconi anemia in Brazilian patients
AU - Pilonetto, Daniela V.
AU - Pereira, Noemi F.
AU - Bonfim, Carmem M. S.
AU - Ribeiro, Lisandro L.
AU - Bitencourt, Marco A.
AU - Kerkhoven, Lianne
AU - Floor, Karijn
AU - Ameziane, Najim
AU - Joenje, Hans
AU - Gille, Johan J. P.
AU - Pasquini, Ricardo
PY - 2017
Y1 - 2017
N2 - BACKGROUND: Fanconi anemia (FA) is a predominantly autosomal recessive disease with wide genetic heterogeneity resulting from mutations in several DNA repair pathway genes. To date, 21 genetic subtypes have been identified. We aimed to identify the FA genetic subtypes in the Brazilian population and to develop a strategy for molecular diagnosis applicable to routine clinical use.METHODS: We screened 255 patients from Hospital de Clínicas, Universidade Federal do Paraná for 11 common FA gene mutations. Further analysis by multiplex ligation-dependent probe amplification (MLPA) for FANCA and Sanger sequencing of all coding exons of FANCA, -C, and -G was performed in cases who harbored a single gene mutation.RESULTS: We identified biallelic mutations in 128/255 patients (50.2%): 89, 11, and 28 carried FANCA,FANCC, and FANCG mutations, respectively. Of these, 71 harbored homozygous mutations, whereas 57 had compound heterozygous mutations. In 4/57 heterozygous patients, both mutations were identified by the initial screening, in 51/57 additional analyses was required for classification, and in 2/57 the second mutation remained unidentified. We found 52 different mutations of which 22 were novel.CONCLUSION: The proposed method allowed genetic subtyping of 126/255 (49.4%) patients at a significantly reduced time and cost, which makes molecular diagnosis of FA Brazilian patients feasible.
AB - BACKGROUND: Fanconi anemia (FA) is a predominantly autosomal recessive disease with wide genetic heterogeneity resulting from mutations in several DNA repair pathway genes. To date, 21 genetic subtypes have been identified. We aimed to identify the FA genetic subtypes in the Brazilian population and to develop a strategy for molecular diagnosis applicable to routine clinical use.METHODS: We screened 255 patients from Hospital de Clínicas, Universidade Federal do Paraná for 11 common FA gene mutations. Further analysis by multiplex ligation-dependent probe amplification (MLPA) for FANCA and Sanger sequencing of all coding exons of FANCA, -C, and -G was performed in cases who harbored a single gene mutation.RESULTS: We identified biallelic mutations in 128/255 patients (50.2%): 89, 11, and 28 carried FANCA,FANCC, and FANCG mutations, respectively. Of these, 71 harbored homozygous mutations, whereas 57 had compound heterozygous mutations. In 4/57 heterozygous patients, both mutations were identified by the initial screening, in 51/57 additional analyses was required for classification, and in 2/57 the second mutation remained unidentified. We found 52 different mutations of which 22 were novel.CONCLUSION: The proposed method allowed genetic subtyping of 126/255 (49.4%) patients at a significantly reduced time and cost, which makes molecular diagnosis of FA Brazilian patients feasible.
UR - https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85041401212&origin=inward
UR - https://www.ncbi.nlm.nih.gov/pubmed/28717661
U2 - https://doi.org/10.1002/mgg3.293
DO - https://doi.org/10.1002/mgg3.293
M3 - Article
C2 - 28717661
SN - 2324-9269
VL - 5
SP - 360
EP - 372
JO - Molecular genetics and genomic medicine
JF - Molecular genetics and genomic medicine
IS - 4
ER -