High-throughput genotyping with infrared fluorescence allele specific hybridization (iFLASH): a simple, reliable and low-cost alternative.
OBJECTIVE
To develop and validate a novel genotyping approach, named infrared Fluorescence Allele Specific Hybridization (iFLASH), which combines the principles of allele specific oligonucleotide (ASO) hybridization with the advanced possibilities of infrared imaging.


METHODS
As an example, we genotyped the 55L > M and the 192Q > R common genetic variants of the paraoxonase-1 gene in 92 DNA samples using the iFLASH technique, and validated the outcomes with the restriction fragment length polymorphism (RFLP) and TAQman genotyping assays.


RESULTS
There was a 100 percent agreement in genotype outcome among the three methods.


CONCLUSIONS
Although we found complete unity in genotype outcome, the iFLASH assay has essential advantages over the RFLP and TAQman genotyping assays. First, the iFLASH technique is capable of handling up to 1536 samples per assay, which makes it a suitable technique for high-throughput genotyping. Secondly, because the costs per assay are lower, high-throughput genotyping with iFLASH is affordable.
van Himbergen
Thomas M
TM
Research Laboratory of the Department of Clinical Chemistry, University Medical Center Utrecht, Utrecht, The Netherlands. T.M.vanHimbergen@azu.nl
Voorbij
Hieronymus A M
HA
Barendrecht
Arjan D
AD
van Rijn
Bas B
BB
Brambilla
Roberta
R
van Tits
Lambertus J H
LJ
Roest
Mark
M
D000483:Alleles; D043303:Aryldialkylphosphatase / Q000235:genetics; D014644:Genetic Variation; D005838:Genotype; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000191:economics* / Q000379:methods*; D007259:Infrared Rays; D012150:Polymorphism, Restriction Fragment Length; D015203:Reproducibility of Results; D012815:Signal Processing, Computer-Assisted
D043303:Aryldialkylphosphatase; C468406:PON1 protein, human