Spatially addressable bead-based biosensor for rapid detection of beta-thalassemia mutations.
A simple glass-polymer bead-based biosensor was validated for the detection of beta-thalassemia mutations. Different bead types, each carrying allele-specific probes targeting a particular mutation on the beta-globin gene, were immobilized and distinguished on the chip based on their spatial addresses. Genomic DNA samples carrying various single nucleotide transitions and transversions in the beta-globin gene were subjected to polymerase chain reaction and asymmetric amplification in the presence of Cy3-labeled primers, followed by hybridization onto the chip and detection under an epifluorescent microscope. Mutations that were heterozygous or homozygous were easily detected on the device based on the signal intensity difference (or similarity) between the wildtype and mutant probes. This device successfully detected all six common beta-globin gene mutations within 30 min. The number of targeted mutations on this chip can be easily expandable through the introduction of additional probe sets.
Ng
Johnson Kian-Kok
JK
Department of Paediatrics, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
Wang
Wen
W
Liu
Wen-Tso
WT
Chong
Samuel S
SS
D000483:Alleles; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D004252:DNA Mutational Analysis / Q000379:methods*; D006801:Humans; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D016133:Polymerase Chain Reaction; D011108:Polymers / Q000737:chemistry; D055544:beta-Globins / Q000235:genetics*; D017086:beta-Thalassemia / Q000175:diagnosis / Q000235:genetics*
D011108:Polymers; D055544:beta-Globins