Objectives:
(i)Subtyping of Influenza A: Toehold switch can subtype Influenza A by complementary pairing between detecting region with of hemagglutimin gene and neuramindase gene.
(ii)On-site detection:A paper-based detection system is portable and readily available for inspections in chicken farm and country border.
(iii)Intuitive readout:The interpretation of result is intuitive and easy to layman by simply reading different combination of reporter signals on test paper (Fig 1).
(iv)Semi-automatic toehold switch design: By linking to Influenza A sequence database, and embedding Vienna RNA (8) in our program, we can minimize the manpower and time on toehold switch design, while giving reliable results.
(v)Standardized toehold switch cloning: With a cloning standard, a new switch can be made by synthesizing the detection sequence and inserting it into standard vector.