Validation of Fanconi anemia complementation Group A assignment using molecular analysis.
OBJECTIVE
Fanconi anemia is a genetically heterogeneous chromosomal breakage disorder exhibiting a high degree of clinical variability. Clinical diagnoses are confirmed by testing patient cells for increased sensitivity to crosslinking agents. Fanconi anemia complementation group assignment, essential for efficient molecular diagnosis of the disease, had not been validated for clinical application before this study. The purpose of this study was (1) confirmation of the accuracy of Fanconi anemia complementation group assignment to Group A (FANCA) and (2) development of a rapid mutation detection strategy that ensures the efficient capture of all FANCA mutations.


METHODS
Using fibroblasts from 29 patients, diagnosis of Fanconi anemia and assignment to complementation Group A was made through breakage analysis studies. FANCA coding and flanking sequences were analyzed using denaturing high pressure liquid chromatography, sequencing, and multiplex ligation-dependent probe amplification. Patients in which two mutations were not identified were analyzed by cDNA sequencing. Patients with no mutations were sequenced for mutations in FANCC, G, E, and F.


RESULTS
Of the 56 putative mutant alleles studied, 89% had an identifiable FANCA pathogenic mutation. Eight unique novel mutations were identified.


CONCLUSIONS
Complementation assignment to Group A was validated in a clinical laboratory setting using our FANCA rapid molecular testing strategy.
Department of Molecular and Medical Genetics, Clinical Genetics Laboratories, Oregon Health & Science University, 3181 S.W. Sam Jackson Park Road, Portland, OR 97239, USA.
Nabil N
NN
Moghrabi
Monique A
MA
Johnson
Marvin J
MJ
Yoshitomi
Xiaoman
X
Zhu
Muhsen J
MJ
Al-Dhalimy
Susan B
SB
Olson
Markus
M
Grompe
C Sue
CS
Richards
D000293:Adolescent; D002460:Cell Line; D002648:Child; D020732:Cytogenetic Analysis; D004252:DNA Mutational Analysis / Q000379:methods; D005199:Fanconi Anemia / Q000145:classification / Q000175:diagnosis / Q000235:genetics*; D052217:Fanconi Anemia Complementation Group A Protein / Q000235:genetics*; D005260:Female; D006801:Humans; D008297:Male; D009154:Mutation; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D055815:Young Adult
D052217:Fanconi Anemia Complementation Group A Protein