Patent ID: 8512955

Claim:
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 target capture oligomers and with at least two different amplification oligomer complexes; (i) wherein each of said target capture oligomers and each of said amplification oligomer complexes hybridize to different target nucleic acid sequences, (ii) wherein each of said amplification oligomer complexes comprises a first amplification oligomer member having a first target specific sequence that is directly joined to a second amplification oligomer member having a second target specific sequence; (b) contacting said at least two target capture oligomers in step (a) with a solid support and immobilized probe for performing a target capture; (c) washing said sample after target capture; (d) pre-amplifying said target nucleic acid sequences using said amplification oligomer complexes, under conditions configured to maintain the amplification oligomer complexes, thereby generating first amplification products for each target nucleic acid hybridized by an amplification oligomer complex; (e) splitting said pre-amplified sample into at least two separate secondary target specific amplification reactions; (f) amplifying at least two of said pre-amplified samples generated in step (d) using target specific amplification oligomers that are not joined, thereby generating second amplification products.