Label free detection of nucleic acid amplification

摘要

Provided are methods and devices for label-free detection of nucleic acids that are amplified by polymerase chain reaction. A solution containing the components necessary for a PCR is introduced to a microfluidic amplification chamber and an electric field applied to a confined region in which PCR occurs. PCR product generated in the confined region is detected by measuring an electrical parameter that is, for example, solution impedance. The devices and methods provided herein are used, for example, in assays to detect one or more pathogens or for point-of-care tests. In an aspect, the PCR product is confined to droplets and the assay relates to detecting an electrical parameter of a flowing droplet, thereby detecting PCR product without a label. In an aspect, the PCR occurs in the droplet.

主权项

1. A method of detecting a nucleic acid amplification product in solution, said method comprising:
providing a microfluidic amplification chamber comprising an electrode array on at least one surface of said microfluidic amplification chamber; introducing a solution that may contain a template to be amplified by nucleic acid amplification in said microfluidic amplification chamber; performing a nucleic acid amplification on said solution, wherein said nucleic acid amplification is performed in a confined region or said nucleic acid amplification product is contained in a confined region, to generate an amplified nucleic acid amplification product in solution, wherein said confined region has a volume selected from a range that is greater than or equal to 0.5 pL and less than or equal to 1 nL; applying an electric potential to said electrode array to produce an electric field in said confined region; and measuring an electrical parameter of said solution in said confined region, wherein said electrical parameter is solution impedance, thereby detecting said nucleic acid amplification product in solution; wherein said step of introducing the solution comprises: providing a droplet of the solution surrounded by a suspending fluid comprising an ionic liquid, and the droplet corresponds to the confined region; and electrically contacting the droplet with the electrode array; wherein said detection of nucleic acid amplification is label-free.