Cystic fibrosis carrier screening: validation of a novel method using BeadChip technology.
OBJECTIVE
To validate a novel BeadChip assay system for cystic fibrosis (CF) mutation testing using the panel of 25 ACMG recommended mutations and D1152H.


METHODS
DNA from 519 individuals originally tested for CF mutation status by allele specific oligonucleotide hybridization (ASOH) were blindly analyzed by the BeadChip assay and the results were compared. The elongation mediated multiplexed analysis of polymorphisms (eMAP) protocol, which combines multiplex amplification of genomic DNA and multiplex detection of mutations on color-coded bead arrays, was used to analyze 26 CF mutations in two separate groups.


RESULTS
The system accurately distinguished the 26 CF genotypes and had 100% concordance with the ASOH technique with an assay failure rate of 1.7%. Benign variants of exon 10 codons 506, 507, and 508 did not interfere with mutation identification and reflex testing for the 5/7/9T IVS8 polymorphism was performed on a separate array.


CONCLUSIONS
The BeadChip assay system provided accurate and rapid identification of the ACMG recommended CF mutations.
Edelmann
Lisa
L
Department of Human Genetics, Mount Sinai School of Medicine, New York, NY 10029, USA.
Hashmi
Ghazala
G
Song
Yuhua
Y
Han
Yi
Y
Kornreich
Ruth
R
Desnick
Robert J
RJ
D003550:Cystic Fibrosis / Q000175:diagnosis* / Q000235:genetics; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics; D004252:DNA Mutational Analysis; D015342:DNA Probes; D006580:Genetic Carrier Screening; D005820:Genetic Testing; D006801:Humans; D009154:Mutation; D009693:Nucleic Acid Hybridization
C505032:CFTR protein, human; D015342:DNA Probes; D019005:Cystic Fibrosis Transmembrane Conductance Regulator