Patent ID: 9738941
Date: 2017-08-22
CPC Classifications: C12Q

Claim:
1. A method of screening a sample for a multiplicity of respiratory pathogens to detect a particular pathogen, the method comprising: (a) isolating nucleic acid from the sample, which nucleic acid putatively comprises nucleic acid from one or more respiratory pathogens; (b) subjecting the nucleic acid to solid phase amplification in the presence of two or more combinations of aqueous primer pairs and corresponding oligonucleotide probes immobilized to a bead in a beadset, wherein the aqueous primer pairs comprise forward primers selected from the group consisting of SEQ ID NOs:1 to 16, 33 and 35 and corresponding reverse primers selected from the group consisting of SEQ ID NOs:17 to 32, 34 and 36, respectively, and the corresponding oligonucleotide probes are selected from the group consisting of SEQ ID NOs:37 to 54, or the primer pairs and corresponding probes are selected from two or more of the sequences listed in Table 8 (SEQ ID NOs:74 to 141); wherein an aqueous primer pair directs the amplification of a region of nucleic acid from a respiratory pathogen, the number of primer pairs being selected on the basis of the number of pathogens desired to be screened and wherein at least one member of the aqueous primer pair comprises a first optically detectable label that is incorporated into a resulting amplicon following amplification; wherein the amplicon is captured by hybridizing to an oligonucleotide probe that is complementary to a region of the amplicon and immobilized to a bead in a beadset, the beadset having subsets of beads, each subset being homogenous with respect to bead size and, optionally, intensity of a second optically detectable label, thereby creating a heterogeneous beadset based on size and/or second detectable label intensity and wherein the number of subsets corresponds to the number of respiratory pathogens to be screened; (c) determining to which of the beads an amplicon has bound on the basis of the intensity of the first detectable label and, where amplicons are bound to multiple subsets of beads, distinguishing between the multiple subsets of beads on the basis of bead size and, optionally, on the basis of second optically detectable label intensity; wherein binding of an amplicon to a particular subset of beads is indicative of the presence of a particular respiratory pathogen in the sample.