Real-time multiplex analysis of four beta-thalassemia mutations employing surface plasmon resonance and biosensor technology.
In this paper, biospecific interaction analysis (BIA) employing surface plasmon resonance (SPR) and biosensor technologies was applied to the analysis of multiple mutations of the human beta-globin gene. To this aim, large target polymerase chain reaction (PCR) products were immobilized on sensor chips and then probes detecting beta degrees 39 (C>T), beta degrees IVSI-1 (G>A), beta(+)IVSI-6 (T>C) and beta(+)IVSI-110 (G>A) thalassemia mutations were sequentially injected. In this study, a total of ten normal and seven heterozygous subjects, and six homozygous patients were considered. The results obtained allow to conclude that discrimination between normal subjects, heterozygous, and homozygous patients is readily achieved for all the four mutations by PCR amplification of genomic DNA containing all the regions corresponding to the same mutations, immobilization of the same PCR products, and hybridization. To our knowledge the procedure described here is the first reported on the use of SPR-based BIA and biosensor technology for multiple detections of point mutations.
Feriotto
Giordana
G
Laboratory for the Development of Pharmacological and Pharmacogenomic Therapy of Thalassemia, Biotechnology Center, Ferrara University, Ferrara, Italy.
Breveglieri
Giulia
G
Finotti
Alessia
A
Gardenghi
Sara
S
Gambari
Roberto
R
D001483:Base Sequence; D015374:Biosensing Techniques; D004247:DNA / Q000737:chemistry / Q000235:genetics; D004252:DNA Mutational Analysis / Q000379:methods*; D015342:DNA Probes; D005914:Globins / Q000235:genetics*; D006579:Heterozygote; D006720:Homozygote; D006801:Humans; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D020349:Surface Plasmon Resonance; D017086:beta-Thalassemia / Q000175:diagnosis / Q000235:genetics*
D015342:DNA Probes; D005914:Globins; D004247:DNA