Surface plasmon resonance and biosensor technology for real-time molecular diagnosis of beta o 39 thalassemia mutation.
BACKGROUND
Biospecific interaction analysis (BIA) employing surface plasmon resonance (SPR) and biosensor technologies is of interest in clinical genetics. However, few data are available on its use in hereditary diseases caused by genetic mutations.


OBJECTIVE
The primary aim of this study was the refinement of BIA technology for use in identifying the beta o 39 mutation of the beta-globin gene, a mutation which causes a common type of beta o thalassemia.


METHODS
Target-biotinylated PCR products were immobilized on streptavidin-coated sensor chips and diagnosed using SPR-based BIA performed by injecting specific oligonucleotide probes into the sensor chip.


RESULTS
We demonstrated that the beta o 39 mutation can be easily and reproducibly identified during the association phase.


CONCLUSIONS
This should be considered a pilot study demonstrating the ability of SPR-based BIA to detect point mutations in the beta-globin gene by real-time monitoring of hybridization between oligonucleotide probes and target-biotinylated PCR products generated from genomic DNA from normal, heterozygous individuals and homozygous beta o thalassemia patients.
Feriotto
Giordana
G
Laboratory for the Development of Pharmacological and Pharmacogenomic Treatment of Thalassemia, Biotechnology Center, Ferrara University, Ferrara, Italy.
Breveglieri
Giulia
G
Gardenghi
Sara
S
Carandina
Gianni
G
Gambari
Roberto
R
D001483:Base Sequence; D015374:Biosensing Techniques / Q000379:methods*; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000235:genetics; D005914:Globins / Q000235:genetics*; D006579:Heterozygote; D006720:Homozygote; D006801:Humans; D009154:Mutation; D009690:Nucleic Acid Conformation; D015345:Oligonucleotide Probes / Q000235:genetics; D010865:Pilot Projects; D016133:Polymerase Chain Reaction; D020349:Surface Plasmon Resonance / Q000379:methods*; D017086:beta-Thalassemia / Q000175:diagnosis* / Q000235:genetics*
D004277:DNA, Single-Stranded; D015345:Oligonucleotide Probes; D005914:Globins