Patent ID: 9169512
Filing Date: 2015-10-27
Classification: C12Q

Abstract:
1. A method for simultaneously amplifying at least two different target nucleic acid sequences contained in a sample comprising the steps of: (a) contacting a sample with at least two different amplification oligomer complexes that hybridize to different target nucleic acid sequences and with at least two target capture oligomers that hybridize to different target nucleic acid sequences, wherein each of said amplification oligomer complexes comprises a first amplification oligomer member that is directly joined to a second amplification oligomer member; (b) providing conditions for hybridizing each of said two amplification oligomer complexes and each of said target capture oligomers to their target nucleic acids present in said sample, and for hybridizing a capture region of said target capture oligomers to a solid support structure; (c) performing a wash step to remove all unhybridized amplification oligomer complexes; (d) pre-amplifying said target nucleic acid sequences using said amplification oligomer complexes in an isothermal amplification reaction, thereby generating a first amplification product for each target nucleic acid hybridized by an amplification oligomer complex; (e) splitting said pre-amplified sample from step (d) into at least two separate secondary target specific amplification reactions; (f) amplifying said pre-amplified sample in each of said secondary target specific amplification reactions using target specific amplification oligomers, thereby generating second amplification products; and (g) detecting said second amplification products from at least one of said secondary target specific amplification reactions, wherein said detecting step determines the presence of at least one of said at least two different target nucleic acids.