Isothermal amplification using modified primers for rapid electrochemical analysis of coeliac disease associated DQB1*02 HLA allele.
- Sallam Al-Madhagi,
- Hamdi Joda,
- Miriam Jauset-Rubio,
- Mayreli Ortiz,
- Ioanis Katakis,
- Ciara K O'Sullivan
Anal Biochem 2018 Jun 16. Epub 2018 Jun 16.
Interfibio Research Group, Department of Chemical Engineering, Universitat Rovira i Virgili, Avinguda Països Catalans 26, 43007, Tarragona, Spain; ICREA, Passeig Lluís Companys 23, 08010, Barcelona, Spain. Electronic address:
DNA biosensors are attractive tools for genetic analysis as there is an increasing need for rapid and low-cost DNA analysis, primarily driven by applications in personalized pharmacogenomics, clinical diagnostics, rapid pathogen detection, food traceability and forensics. A rapid electrochemical genosensor detection methodology exploiting a combination of modified primers for solution-phase isothermal amplification, followed by rapid detection via hybridization on gold electrodes is reported. Modified reverse primers, exploiting a C18 spacer between the primer-binding site and an engineered single stranded tail, are used in a recombinase polymerase amplification reaction to produce an amplicon with a central duplex flanked by two single stranded tails. Read More