Journal
ANALYTICAL BIOCHEMISTRY
Volume 474, Issue -, Pages 66-68Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2014.11.021
Keywords
Hybridization refractory mutation system; Single base mismatch; Electrochemical genosensor; DQA1*05:05 allele
Funding
- [FP7-2007-ICT-1-216031]
- ICREA Funding Source: Custom
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We report on a simple approach to enhance solid-phase hybridization-based single base mismatch discrimination at high ionic strength based on the deliberate insertion of a natural DNA base mismatch in the surface-tethered probe. A large drop in hybridization signal of single base mismatched alleles using the designed probe as compared with the conventional probe, from 80% to less than 25% of the signal obtained with the fully complementary, non-mutation-containing sequence, when using colorimetric detection was further improved to 20% when using electrochemical detection, attributable to a difference of spacing of immobilized probes. Finally, the designed probe was used for the electrochemical detection of the DQA1*05:05 allele amplified from real human blood samples. (C) 2014 Elsevier Inc. All rights reserved.
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