High-throughput mutational screening for beta-thalassemia by single-nucleotide extension.
In this work a high-throughput method based on the single-nucleotide extension (SNE) reaction and multicolour detection in a DNA sequencer was developed to screen for eight mutations in the human beta-globin gene: IVSI.110, cd39, IVSI.1, IVSI.6, IVSII.745, HbC, HbS and cd6. The method has been validated on a large number of samples for the two most common mutations causing beta-thalassemia in the Mediterranean area (IVSI.110 and cd39). The development of a high-throughput, fast and reliable method to assay beta-thalassemia mutations represents a significant improvement in molecular diagnosis of this disease. The multicolour detection and the use of multiple injections further enhances the throughput of mutational screening by the DNA sequencer and facilitates automated genotyping for routine molecular diagnostics.
Genomic Unit for the Diagnosis of Human Pathologies, San Raffaele Scientific Institute, Milano, Italy.
Silvia
S
Galbiati
Marcella
M
Chiari
Micol
M
Macellari
Maurizio
M
Ferrari
Laura
L
Cremonesi
Marina
M
Cretich
D001483:Base Sequence / Q000235:genetics; D004252:DNA Mutational Analysis / Q000379:methods*; D019075:Electrophoresis, Capillary / Q000379:methods; D020022:Genetic Predisposition to Disease; D005820:Genetic Testing / Q000379:methods*; D005914:Globins / Q000235:genetics*; D006801:Humans; D009154:Mutation; D009711:Nucleotides / Q000378:metabolism*; D016133:Polymerase Chain Reaction / Q000379:methods; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D017086:beta-Thalassemia / Q000175:diagnosis* / Q000235:genetics*
D009711:Nucleotides; D005914:Globins